Amnion Membrane Research Articles

The collection and application of autologous amniotic fluid to cesarean delivery closure – a feasibility study

Amniotic fluid and amnion membranes have been used in surgery specialties to improve wound healing. The objective was to determine if amniotic fluid could be collected at cesarean delivery and then reapplied to the layers of the closure using the CeaLogic Specimen Collection and Ratio Applicator Kit. Autologous amniotic fluid from 20 subjects was collected at cesarean delivery using the collection kit, and then transferred to the applicator kit, mixed with calcium chloride solution, and applied directly to each of the surgical repair layers during closure. The Modified Hollander Cosmesis Score was used to assess wound appearance from photographs of the incision taken in the postpartum setting. The median (IQR) Modified Hollander Cosmesis Score (range 0-best, to 6-worst) at the one week and four-week postpartum visits was 0 (0,1) and 0 (0,2), respectively. There were no wound complications nor surgical site infections. The CeaLogic Specimen Collection and Ratio Applicator Kits can be used to collect and reapply autologous amniotic fluid at cesarean delivery. Future studies can determine if the application of autologous amniotic fluid to cesarean delivery closure can improve cosmesis and wound healing, as well as decrease the risk for the development of intraabdominal adhesions.

Analysis of morphology, cytotoxicity, and water content characteristics of freeze-dried amnion membrane from human and bovine

Placenta tissue has biological advantages, including anti-inflammatory, anti-bacterial, anti-fibrotic formation, and immunomodulatory properties. The amnion membrane (AM) is an inner side membrane of the placenta that faces the fetus. The main sources of amnion are humans and animals, with bovine being one of the significant sources. The aim of this study was to analyze the morphology, cytotoxicity, and water content characteristic of freeze‑dried amnion membrane (FD‑AM) from humans and bovines to measure the safety and compatibility of bovine FD-AM as an alternative to human FD-AM. This study is an observational cross-sectional study. Samples were divided into two groups: human FD-AM and bovine FD-AM groups. Both groups were examined for morphology characteristics by scanning electron microscopy (SEM), cytotoxicity by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) analysis, and water content by drying through moisture analyzer device. The morphology characteristics of bovine FD-AM and human FD-AM, as observed through SEM, showed similar results of a smooth, flat surface with no cavity and were well dehydrated. MTT assay analysis on both groups demonstrated cytocompatibility with cell viability exceeding 70% in the control group. However, human FD-AM showed a higher number of viable cells (0.19±0.01) compared to bovine FD-AM (0.12±0.03), with a statistically significant difference (p<0.05). The water content analysis revealed that both groups met the standard, with levels below 10%. While bovine FD-AM (7.19±0.45%) had slightly higher water content than human FD-AM (6.79±1.0%), the difference was not significant (p>0.05). Both human FD-AM and bovine FD-AM showed good results in morphology, cytotoxicity, and water content characteristics and compatibility. In conclusion, bovine FD-AM might be considered as an alternative to human FD-AM.

The prevalence of inflammatory process in placentas from premature births in different gestational periods

Premature birth (PB) is the birth in less than 37 weeks, which can be spontaneous and iatrogenic, its frequency reaches 10% of all births. The pathogenesis of premature labor is based on the inflammatory reaction of the chorion, amnion and decidual membrane, but the origin of the inflammatory reaction is different depending on the gestational age.The objective: to study the prevalence of the inflammatory response in placenta samples from very early preterm, early preterm and term births.Materials and methods. Placenta samples from 203 parturient women who gave birth in the gestational period of 22–27 weeks – very early PB (I group – 101 parturient women with premature rupture of membranes (PROM), II group – 102 parturient women with the onset of labor with intact membranes) and 202 parturient women were examined with deliveries in the period of 28–34 weeks – early PB (III group – 100 women in labor with PROM and IV group – 102 women in labor with the onset of labor with intact fetal membranes) and 60 women in labor with term deliveries.Immunohistochemical detection of CD8, CD3 and CD14 antigens in villi was used to assess the prevalence of the inflammatory process and compare it between groups.Results. In 56.7% of placentas from term deliveries no markers of villousitis were detected, a mild form of villousitis was found in another third of cases. In placentas of woman with the onset of labor between the 28th and 34th weeks with intact fetal membranes there were signs of villousitis in 54.9%, and its mild manifestations in 33.3%. In the case of premature rupture of the membranes, the prevalence of villousitis reached 65%, its severe forms were more often observed.On the other hand, in the groups of very early PB only 14.9 and 15.7% of placentas had no signs of vilouisitis, one in five placentas from these groups had severe focal or diffuse villousitis. In the group of early PB and intact amniotic membranes, one third of the placentas demonstrated an isolated CD3 lymphocyte marker, no antigen was detected in 47.1% of placentas, and only in every tenth placenta antigens of macrophages of fetal origin were determined. A similar distribution was found in placentas from early PB with PROM.In placentas from very early PB the highest frequency of CD14 detection was found – every second placenta contained this marker in isolation and in 22.5% – in combination with maternal lymphocyte markers. In the group of very early PB on the background of PROM placentas with a positive CD14 test dominated – 31.7% isolated and 45.5% in combination with CD3.Conclusions. 1. In placentas from premature births (PB) at 28–34 weeks, villousitis was detected in 54.9% of cases against the background of intact amniotic membranes and in 65.0% – on the background of premature rupture of the membranes, from very early PBs – in 85.1% and 83.3%, respectively. Placentas from deliveries with premature rupture of the fetal membranes more often had signs of severe villousitis – in 21% of cases at 28–34 weeks and in 20.6% at 22–27 weeks.2. In placentas from early PB, macrophage antigen of fetal origin was detected in 22% of cases in the cases of premature rupture of the fetal membranes and in 21.6% of cases of childbirth with intact membranes; these samples were dominated by infiltration by lymphocytes of maternal origin. On the other hand, placentas from very early PB and ruptured fetal membranes in 77.2% of cases contained antigen of fetal macrophages and in 71.6% of cases of onset of labor with intact membranes.Such results indicate the fetal origin of the inflammatory process in very early PB.

Decellularized Amnion Membrane Triggers Macrophage Polarization for Desired Host Immune Response.

Appropriate regulation of immunomodulatory responses, particularly acute inflammation involving macrophages, is crucial for the desired functionality of implants. Decellularized amnion membrane (DAM) is produced by removing cellular components and antigenicity, expected to reduce immunogenicity and the risk of inflammation. Despite the potential of DAM as biomaterial implants, few studies have investigated its specific effects on immunomodulation. Here, it is demonstrated that DAM can regulate macrophage-driven inflammatory response and potential mechanisms are investigated. In vitro results show that DAM significantly inhibits M1 polarization in LPS-induced macrophages by inhibiting Toll-like receptors (TLR) signaling pathway and TNF signaling pathway and promotes macrophage M2 polarization. Physical signals from the 3D micro-structure and the active protein, DCN, binding to key targets may play roles in the process. In the subcutaneous implant model in rats, DAM inhibits the persistence of inflammation and fibrous capsule formation, while promoting M2 macrophage polarization, thereby facilitating tissue regeneration. This study provides insights into DAM's effect and potential mechanisms on the balance of M1/M2 macrophage polarization in vitro and vivo, emphasizing the immunomodulation of ECM-based materials as promising implants.

A dynamic flow fetal membrane organ-on-a-chip system for modeling the effects of amniotic fluid motion.

Fetal membrane (amniochorion), the innermost lining of the intrauterine cavity, surround the fetus and enclose amniotic fluid. Unlike unidirectional blood flow, amniotic fluid subtly rocks back and forth, and thus, the innermost amnion epithelial cells are continuously exposed to low levels of shear stress from fluid undulation. Here, we tested the impact of fluid motion on amnion epithelial cells (AECs) as a bearer of force impact and their potential vulnerability to cytopathologic changes that can destabilize fetal membrane functions. A previously developed amnion membrane (AM) organ-on-chip (OOC) was utilized but with dynamic flow to culture human fetal amnion membrane cells. The applied flow was modulated to perfuse culture media back and forth for 48h to mimic fluid motion. A static culture condition was used as a negative control, and oxidative stress (OS) condition was used as a positive control representing pathophysiological changes. The impacts of fluidic motion were evaluated by measuring cell viability, cellular transition, and inflammation. Additionally, scanning electron microscopy (SEM) imaging was performed to observe microvilli formation. The results show that regardless of the applied flow rate, AECs and AMCs maintained their viability, morphology, innate meta-state, and low production of pro-inflammatory cytokines. E-cadherin expression and microvilli formation in the AECs were upregulated in a flow rate-dependent fashion; however, this did not impact cellular morphology or cellular transition or inflammation. OS treatment induced a mesenchymal morphology, significantly higher vimentin to cytokeratin 18 (CK-18) ratio, and pro-inflammatory cytokine production in AECs, whereas AMCs did not respond in any significant manner. Fluid motion and shear stress, if any, did not impact AEC cell function and did not cause inflammation. Thus, when using an amnion membrane OOC model, the inclusion of a dynamic flow environment is not necessary to mimic in utero physiologic cellular conditions of an amnion membrane.

Mayer–Rokitansky–Küster–Hauser syndrome managed with McIndoe’s vaginoplasty: a case series and literature review

Introduction: Mayer–Rokitansky–Küster–Hauser (MRKH) syndrome is a congenital anomaly characterized by the absence of the uterus and the upper two-thirds of the vagina. It is a rare congenital anomaly with an incidence of 1 in 5000 female live births. Case series: The authors describe three cases of females presenting with primary amenorrhoea who were diagnosed with MRKH syndrome. The patients were managed with McIndoe’s vaginoplasty with neovagina creation with an amnion graft. Discussion: Management of MRKH syndrome involves vaginoplasty with neovagina creation. The approach to neovagina creation can be done surgically or non-surgically. Non-surgical creation of the vaginal cavity involves serial use of vaginal dilators, while there are several ways for surgical creation of neovagina. The modified Abbe-McIndoe procedure using amnion to create neovagina is a minimally invasive, rapid, and simple procedure with no risk of immune rejection because the amnion membrane lacks histocompatibility antigens. In addition, the graft is also readily available, storable, and inexpensive. Conclusion: Diagnosis of MRKH syndrome can be made when a young female with primary amenorrhoea and normal secondary sexual characteristics has agenesis of the uterus, and upper two-thirds of the vagina revealed on ultrasonography or magnetic resonance imaging. The patient can be offered treatment with vaginoplasty with neovagina creation.

The bioengineering application of hyaluronic acid in tissue regeneration and repair

The multifaceted role of hyaluronic acid (HA) across diverse biomedical disciplines underscores its versatility in tissue regeneration and repair. HA hydrogels employ different crosslinking including chemical (chitosan, collagen), photo- initiation (riboflavin, LAP), enzymatic (HRP/H2O2), and physical interactions (hydrogen bonds, metal coordination). In biophysics and biochemistry, HA's signaling pathways, primarily through CD44 and RHAMM receptors, modulate cell behavior (cell migration; internalization of HA), inflammation, and wound healing. Particularly, smaller HA fragments stimulate inflammatory responses through toll-like receptors, impacting macrophages and cytokine expression. HA's implications in oncology highlight its involvement in tumor progression, metastasis, and treatment. Elevated HA in tumor stroma impacts apoptosis resistance and promotes tumor growth, presenting potential therapeutic targets to halt tumor progression. In orthopedics, HA's presence in synovial fluid aids in osteoarthritis management, as its supplementation alleviates pain, enhances synovial fluid's viscoelastic properties, and promotes cartilage integrity. In ophthalmology, HA's application in dry eye syndrome addresses symptoms by moisturizing the eyes, replenishing tear film deficiencies, and facilitating wound healing. Intravitreal injections and hydrogel-based systems offer versatile approaches for drug delivery and vitreous humor replacement. For skin regeneration and wound healing, HA hydrogel dressings exhibit exceptional properties by promoting moist wound healing and facilitating tissue repair. Integration of advanced regenerative tools like stem cells and solubilized amnion membranes into HA-based systems accelerates wound closure and tissue recovery. Overall, HA's unique properties and interactions render it a promising candidate across diverse biomedical domains, showcasing immense potentials in tissue regeneration and therapeutic interventions. Nevertheless, many detailed cellular and molecular mechanisms of HA and its applications remain unexplored and warrant further investigation.

A Dynamic Flow Fetal Membrane Organ-on-a-Chip System for Modeling the Effects of Amniotic Fluid Motion.

Fetal membrane(amniochorion), the innermost lining of the intrauterine cavity, surround the fetus and enclose amniotic fluid. Unlike unidirectional blood flow, amniotic fluid subtly rocks back and forth, and thus, the innermost amnion epithelial cells are continuously exposed to low levels of shear stress from fluid undulation. Here, we tested the impact of fluid motion on amnion epithelial cells (AECs) as a bearer of force impact and their potential vulnerability to cytopathologic changes that can destabilize fetal membrane functions. An amnion membrane (AM) organ-on-chip (OOC) was utilized to culture human fetal amnion membrane cells. The applied flow was modulated to perfuse culture media back and forth for 48 hours flow culture to mimic fluid motion. Static culture condition was used as a negative control, and oxidative stress (OS) condition was used as a positive control for pathophysiological changes. The impacts of fluidic motion were evaluated by measuring cell viability, cellular transition, and inflammation. Additionally, scanning electron microscopy (SEM) imaging was performed to observe microvilli formation. The results show that regardless of the applied flow rate, AECs and AMCs maintained their viability, morphology, innate meta-state, and low production of pro-inflammatory cytokines. E-cadherin expression and microvilli formation in the AECs were upregulated in a flow rate-dependent fashion; however, this did not impact cellular morphology or cellular transition or inflammation. OS treatment induced a mesenchymal morphology, significantly higher vimentin to CK-18 ratio, and pro-inflammatory cytokine production in AECs, whereas AMCs did not respond in any significant manner. Fluid motion and shear stress, if any, did not impact AEC cell function and did not cause inflammation. Thus, when using an amnion membrane OOC model, the inclusion of a flow culture environment is not necessary to mimic any in utero physiologic cellular conditions of fetal membrane-derived cells.

Tocolytic effectiveness of nifedipine in management of preterm labor: A tertiary center experience

Objective: This study aimed to compare the efficacy of nifedipine use in our clinic for treatment of preterm labor with diferent effacement degrees, and to compare our results with the literature. Methods: 440 singleton pregnant women with intact amnionic membrane pregnant women in their 23 and 36 weeks were retrospectively evaluated. Because of different criterias defining preterm labor in the literature, patients were divided into two groups as 4 or over contractions in 20 minutes and cervical opening at 2 cm and/or above and/or effachment at 80% or above (Group A, n=230) and 4 or over contractions in 20 minutes and cervical opening below 2 min and effechment below 80% (Group B, n=210). Descriptive statistics were conducted using chi-square and Mann Whitney U test, and statistically significance was p <0.05. Results: Demographics, reproductive history and pregnancy weeks of the groups were similar. The average time between the start of tocolysis and the birth was 4 times higer in Group B (0.1-99.2, mean 28.4) than that of Group A (0.1-78.9, mean 7.4 days, p<0.001). Delay after tocoloysis at days 1 to 7 was statistically remarkable in Group B (p<0.001, for each). Preterm labor resulting in early birth was more remarkable in Group A in both before 34 weeks and 37 weeks (p<0.001, for each; n=88, 38.3% vs n=36, 17.1% and n=171, 74.3% vs n=98, 46.7%, respectively and n=269/124, 28.2 vs 61.1%). Delay with nifedipine at day 1 (87.5%), day 2 (79.1%), day 3 (74.8%) and day 7 (65.5%) was also compatable with the literature. Conclusions: Nifedipine is an effective tocolytic agent in preterm labor regardless of the effachment degree.

PURION® processed human amnion chorion membrane allografts retain material and biological properties supportive of soft tissue repair.

The reparative properties of amniotic membrane allografts are well-suited for a broad spectrum of specialties. Further enhancement of their utility can be achieved by designing to the needs of each application through the development of novel processing techniques and tissue configurations. As such, this study evaluated the material characteristics and biological properties of two PURION® processed amniotic membrane products, a lyophilized human amnion, intermediate layer, and chorion membrane (LHACM) and a dehydrated human amnion, chorion membrane (DHACM). LHACM is thicker; therefore, its handling properties are ideal for deep, soft tissue deficits; whereas DHACM is more similar to a film-like overlay and may be used for shallow defects or surgical on-lays. Characterization of the similarities and differences between LHACM and DHACM was conducted through a series of in vitro and in vivo studies relevant to the healing cascade. Compositional analysis was performed through histological staining along with assessment of barrier membrane properties through equilibrium dialysis. In vitro cellular response was assessed in fibroblasts and endothelial cells using cell proliferation, migration, and metabolic assays. The in vivo cellular response was assessed in an athymic nude mouse subcutaneous implantation model. The results indicated the PURION® process preserved the native membrane structure, nonviable cells and collagen distributed in the individual layers of both products. Although, LHACM is thicker than DHACM, a similar composition of growth factors, cytokines, chemokines and proteases is retained and consequently elicit comparable in vitro and in vivo cellular responses. In culture, both treatments behaved as potent mitogens, chemoattractants and stimulants, which translated to the promotion of cellular infiltration, neocollagen deposition and angiogenesis in a murine model. PURION® processed LHACM and DHACM differ in physical properties but possess similar in vitro and in vivo activities highlighting the impact of processing method on the versatility of clinical use of amniotic membrane allografts.

Efficacy of Amniotic and Chorionic Membrane in Facial Wound Healing: A Comparative Study.

Background Advancements in regenerative techniques have been utilized in placental amnion and chorion for a variety of purposes. Their ability to regenerate tissues has led to their usage in tissue engineering, wound healing, and other therapeutic applications. This study aims to evaluate and compare the efficacy of amnion and chorion in facial tissue wound healing. Methodology The study was an observational comparative study conducted in the Department of Oral and Maxillofacial Surgery, involving 20 participants divided into two groups (Group I and Group II).Study groups were selected according to the inclusion and exclusion criteria. A dehydrated human amnion/ chorion membrane was applied to the affected site of each group respectively. Its efficacy in wound healing was analyzed in the first, third, seventhday, and secondweek. Statistical analysis was done using SPSS software(IBM Corp., Armonk, NY). Results Patients treated with amnion membrane showed a decrease in wound size and the wound was completely healed by secondweek with mean scores of wound sizes of 0.00whereas the wound remained unhealed by secondweek with mean of 1.70to those treated with chorion membrane. Conclusion Amnion showed superior efficacy in wound healing at two-week intervals when compared to the chorion. Hence, this could be used in regenerative medicine as a graft to induce healing in facial wounds.

Evaluation of Regenerative Efficacy of Amnion and Chorion Membrane in Treatment of Mandibular Molar Furcation Defects: A Clinico-radiographic Study.

Amnion and chorion membranes possess unique inherited biological properties that enhance wound healing and may accelerate periodontal regeneration. The present study aims to evaluate and compare the efficacy of amnion and chorion membranes in the treatment of furcation defects. A total of 20 patients were selected and were randomly allocated to group I and group II with 10 subjects in each group. Amnion and chorion membranes are placental-derived membranes that accelerate regeneration by having natural growth factors with their antimicrobial and inflammation reduction properties. Group I was treated using bone grafting with decalcified freeze-dried bone allograft (DFDBA) and placement of amnion as a membrane for guided tissue regeneration (GTR) whereas group II was treated using bone grafting with DFDBA and placement of chorion as a membrane for GTR. The patients were followed for clinical and radiographic parameters and were evaluated between 3 and 6 months after surgery. In intragroup comparison, a significant difference was evident in both the groups for all the clinical and radiographic parameters within the groups. (p = 0.01) This means both amnion and chorion membranes showed statistically significant regenerative efficacy. In intergroup comparison, the results show that all the clinical parameters and radiographic parameters show no significant difference between the groups. The amnion and chorion membranes had similar regenerative efficacy in combination with DFDBA in patients with buccal degree II furcation defects in mandibular molars. The amnion and chorion membranes have shown significant improvement in clinical and radiographic parameters when used for the treatment of buccal degree II furcation defects in mandibular molars. How to cite this article: Mallapragda S, Gupta R, Gupta S, et al. Evaluation of Regenerative Efficacy of Amnion and Chorion Membrane in Treatment of Mandibular Molar Furcation Defects: A Clinico-radiographic Study. J Contemp Dent Pract 2024;25(2):160-167.

Cost effectiveness analysis for commonly used human cell and tissue products in the management of diabetic foot ulcers

AbstractBackground and AimsThis study considers the cost‐effectiveness of commonly used cellular, acellular, and matrix‑like products (CAMPs) of human origin also known as human cell and tissue products (HCT/Ps) in the management of diabetic foot ulcers.MethodsWe developed a 1‐year economic model assessing six CAMPs [cryopreserved placental membrane with viable cells (vCPM), bioengineered bilayered living cellular construct (BLCC), human fibroblast dermal substitute (hFDS), dehydrated human amnion chorion membrane (dHACM), hypothermically stored amniotic membrane (HSAM) and human amnion membrane allograft (HAMA) which had randomized controlled trial evidence compared with standard of care (SoC). CAMPs were compared indirectly and ranked in order of cost‐effectiveness using SoC as the baseline, from a CMS/Medicare's perspective.ResultsThe mean cost, healed wounds (hw) and QALYs per patient for vCPM is $10,907 (0.914 hw, 0.783 QALYs), for HAMA $11,470 (0.903 hw, 0.780 QALYs), for dHACM $15,862 (0.828 hw, 0.764 QALYs), for BLCC $18,430 (0.816 hw, 0.763 QALYs), for hFDS $19,498 (0.775 hw, 0.757 QALYs), for SoC $19,862 (0.601 hw, 0.732 QALYs) and $24, 214 (0.829, 0.763 QALYs) for HSAM respectively. Over 1 year, vCPM results in cheaper costs overall and better clinical outcomes compared to other CAMPs. Following probabilistic sensitivity analysis, vCPM has a 60%, HAMA 40% probability of being cost‐effective then dHACM, hFDS, BLCC, and lastly HSAM using a $100,000/healed wound or QALY threshold.ConclusionsAll CAMPs were shown to be cost‐effective when compared to SoC in managing DFUs. However, vCPM appears to be the most cost‐effective CAMP over the modelled 52 weeks followed by HAMA, dHACM, hFDS, BLCC, and HSAM. We urge caution in interpreting the results because we currently lack head‐to‐head evidence comparing all these CAMPs and therefore suggest that this analysis be updated when more direct evidence of CAMPs becomes available.

Amnion Membrane Transplantation: A Therapy for Persistent Epithelial Defect as A Result of Neurotrophic Keratitis

Introduction : A persistent epithelial defect (PED) is a failure of re-epithelialization and complete healing of the cornea. The primary goal of PED treatment is to create a favorable environment for corneal wound healing. Amniotic membrane transplants (AMT) become one of the refractory cases treated for PED. This study aims to report AMT as therapy for a PED.
 Case Illustration : A fifty-six-year-old male presents with a chief complaint of redness and pain in the left eye in the last2 weeks. The patient complained of blurred vision, tearing, and foreign body sensations. An ophthalmology examination of the left eye revealed that it was close to face finger counting in visual acuity and had a persistent epithelial defect, corneal edema, descemet fold, and a decrease in corneal sensation. The patient was diagnosed with PED and received autologous serum eye drops, levofloxacin eye drops, vitamin A palmitate eye gel, and acyclovir 400mg twice a day, but did not respond adequately. AMT was planned for the patient. The patient's pain level improved one week after surgery.
 Discussion : The most common symptom of PED is pain. There are various etiological factors for PED. Neurotropic keratitis has emerged as one of the causes of PED. Management for PED is carried out sequentially according to the protocol.
 Conclusion : Amniotic membrane transplantation has become an alternative therapy for PED. The amniotic membrane provides re-epithelialization of the cornea and anti-inflammatory properties. The prognosis for AMT as a therapy for PED is quite good.

A Rare Case of Congenital Corneal Keloids

Introduction : A corneal keloid is a uncommon ophthalmic condition that manifest as proliferation of fibrous tissue in the cornea and presents as an enlarging, white, elevated, well circumscribed corneal lesion. It is a relatively uncommon lesion that typically appears after surgery, injury, or corneal perforation. Pathological examination of the resected specimen leading to a diagnosis of corneal keloid.
 Case Illustration : A 6-year-old girl came with chief complaint was blurry vision in both eyes accompanied with a white opacity in her left eye that was gradually growing. The patient has been diagnosed with congenital glaucoma ever since birth. Nobody of her parents acknowledged having a history of inflammatory, infection, or ocular damage. At first glance, her visual acuity appeared to be light reflex in both eyes. Slit lamp biomicroscopy revealed a pearly white, enhanced opacity with sharp edges and smooth surface in the left eye (figure 1). New vessels were present in the lesion adjacent to the limbus. The patient is undergoing keloids excision, multilayer amnion membrane transplant and fine needle diathermy (figure2).
 Discussion : Corneal keloid can present at any age, present as a glistening white enlarging mass on the cornea and well demarcated from the adjacent normal tissue. Histopathologic examination demonstrates a thickened corneal epithelium overlying stroma and hyalinized collagen (figure 3).
 Conclusion : A corneal keloid should be suspected in cases of enlarging white glistening avascular corneal scars. Biopsy and histopathology confirmed the diagnosis.

Comparative Evaluation of Coronally Advanced Flap with and without Amniotic Membrane in the treatment of Localised Miller’s Class I and Class II Gingival Recession Defects: A Randomised Controlled Trial

Introduction: Gingival recession is the exposure of the root surface of the tooth, characterised by the displacement of the gingival margin apically from the Cementoenamel Junction (CEJ). Various techniques have been suggested for treating recession, such as Free Gingival Graft (FGG), Connective tissue grafts, laterally or Coronally Advanced Flaps (CAF), and Guided Tissue Regeneration (GTR)-based procedures. Different membranes have been used for GTR, one of which is the amnion membrane. Aim: To evaluate root coverage using Coronally Advanced Flap (CAF) with and without amniotic membrane in the treatment of localised gingival recession. Materials and Methods: A randomised controlled trial was conducted at Pandit Deendayal Upadhyay Dental College and Hospital, Solapur, Maharashtra, India from March 2017 to February 2018. In this study, 30 teeth were treated, with 15 teeth undergoing CAF with amniotic membrane (Group IITest Group) and 15 teeth treated with CAF without amniotic membrane (Group I-Control Group). Probing Depth (PD), Clinical Attachment Level (CAL), Relative Attachment Level (RAL), Recession Width (RW), Recession Depth (RD), Width of Keratinised Gingiva (WKG), Width of Attached Gingiva (WAG), and gingival biotype were assessed between baseline and after three months. Statistical analysis was performed using paired t-test, independent t-test, and Mann-Whitney U-Test. Statistical Package for Social Sciences (SPSS) software version 20.0 was used for analysis. The α error of 5% (p-value=0.05) and β error of 20% were taken into consideration. Results: The mean age of the study population was 30.36±4.55 years, with 18 males and 12 females. There was a decrease in RD, RW, PD, and a gain in CAL and RAL in both groups after three months compared to the baseline values. Additionally, there was an increase in WKG, WAG, and a significant change in the thickness of the gingival biotype from thin to thick in both groups at the end of three months. On inter group comparison, the changes in PD (p-value=1.00), RD (p-value=1.00), RW (0.176), CAL (0.664), RAL (1.00), WKG (p-value=0.313) were not statistically significant at the end of three months. WAG was higher in Group II compared to Group I (p-value=0.014*), and the mean root coverage was greater in Group II (15.62%) than in the control group (11.642%) (p-value=0.005) at the end of three months. There was a statistically significant difference (p-value=0.005*) in Group II (15.62%) compared to the control group (11.642%). Conclusion: Amniotic membrane in combination with CAF proves to be a successful option for root coverage in localised recession defects. However, histological evidence and a greater number of future longitudinal studies are necessary to study the efficacy of this approach.

The Effect of MultilayeredElectrospun PLLA Nanofibers Coated with Human Amnion or Bladder ECM Proteins on Epithelialization and Smooth Muscle Regeneration in the Rabbit Bladder.

Nanofibrous scaffolds have attracted much attention in bladder reconstruction approaches due to their excellent mechanical properties. In addition, their biological properties can be improved by combination with biological materials. Taking into account the advantages of nanofibrous scaffolds and decellularized extracellular matrix (dECM) in tissue engineering, scaffolds of poly-L-lactic acid (PLLA) coated with decellularized human amnion membrane (hAM) or sheep bladder (SB)-derived ECM proteins are developed (amECM-coated PLLA and sbECM-coated PLLA, respectively). The bladder regenerative potential of modified electrospun PLLA scaffolds is investigated in rabbits. The presence of ECM proteins is confirmed on the nanofibers' surface. Coating the surface of the PLLA nanofibers improves cell adhesion and proliferation. Histological and immunohistochemical evaluations show that rabbits subjected to cystoplasty with a multilayered PLLA scaffold show de novo formation and maturation of the multilayered urothelial layer. However, smooth muscle bundles (myosin heavy chain [MHC] and α-smooth muscle actin [α-SMA] positive) are detected only in ECM-coated PLLA groups. All groups show no evidence of a diverticulumor fistula in the urinary bladder. These results suggest that the biofunctionalization of electrospun PLLA nanofibers with ECM proteins can be a promising option for bladder tissue engineering. Furthermore, hAM can also replace animal-sourced ECM proteins in bladder tissue regeneration approaches.

EP19.10: Second emergent double McDonald cerclage is useful in patients with amnionic membrane bulging into vagina after the first cervical cerclage

EP19.10: Second emergent double McDonald cerclage is useful in patients with amnionic membrane bulging into vagina after the first cervical cerclage

A Prospective Multicenter Study of a Weekly Application Regimen of Viable Human Amnion Membrane Allograft in the Management of Nonhealing Diabetic Foot Ulcers.

Diabetic foot ulcers (DFUs) pose a significant clinical challenge for providers and patients, and often precede devastating complications such as infection, hospitalization, and amputation. Therefore, advanced treatment options are needed to facilitate the healing of chronic DFUs and improve outcomes in this high-risk population. Cryopreserved viable human amnion membrane allograft (vHAMA) has shown great promise in the treatment of recalcitrant DFUs as a supplement to standard of care (SOC). Placental grafts are rich in extracellular matrix proteins, growth factors, and cytokines, which can induce angiogenesis and dermal fibroblast proliferation, resulting in accelerated healing. In this prospective, multicenter single arm trial, 20 patients with nonhealing DFUs received weekly application of vHAMA, in addition to SOC, for up to 12 weeks. The primary study endpoint was proportion of healed wounds at 12 weeks. Secondary endpoints included proportion of wounds healed at 6 weeks, time to heal, and percentage area wound reduction. Subjects were evaluated for ulcer healing and assessed for adverse events at every treatment visit. At study conclusion, 85% of patients receiving vHAMA healed. Ten wounds healed (50%) by 6 weeks, and 17 wounds (85%) healed by 12 weeks. The mean time to heal was 46.6 days (95% CI: 35.1-58.0), and the average number of vHAMAs used was 5.4 (SD: 3.25). The mean PAR was 86.3% (SD: 40.51). Aseptically processed, cryopreserved vHAMA should be considered as a safe and effective option for DFUs refractory to SOC therapy.

Coronally repositioned flap procedure for gingival root coverage using human derived regenerative membranes - A case series

The aim of this case series was to evaluate the effectiveness of platelet rich fibrin (PRF), Amnion and chorionic membrane for root coverage procedures using a coronally repositioned flap (CRF).Three subjects with Miller’s class I gingival recession were randomly selected for treatment using CRF with PRF, Amnion and Chorion membranes. The clinical parameters evaluated at baseline and 6 months follow up included: probing depth (PD), relative attachment level (RAL), recession height (RH), recession width (RW), width of keratinized tissue (WKT) and Gingival thickness (GT). Within the limits, this case series has shown that the use of human derived regenerative membranes along with CRF resulted in complete root coverage in Miller’s Class I gingival recessions in much lesser time. The rapid healing and regeneration of these membranes stands as an alternative for connective tissue autografts and holds promise for future.Gingival thickness (GT), Relative Attachment level (RAL), Width of Keratinized Tissue (WKT)