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This article explores the potential benefits and challenges of using the amniotic membrane in surgical procedures. Amniotic membranes have been shown to possess anti-inflammatory, anti-scarring, and antimicrobial properties, which can promote wound healing and tissue regeneration. They can be utilized in a variety of surgical settings, including ophthalmology, plastic surgery, and orthopedics. However, the use of amniotic membranes also poses certain limitations and risks, such as the potential for immune reactions and disease transmission. The article reviews the current literature on amniotic membrane usage in surgery and discusses the clinical implications and future research directions. In summary, the amniotic membrane represents a promising alternative to traditional surgical approaches, but its benefits and limitations must be carefully weighed and assessed in each individual case.

In this study, a biological conduit, consisting of an adipocyte-derived mesenchymal stem cell (AdMSCs) sheet and amniotic membrane (AM), was designed for the reconstruction of peripheral nerve defects. To evaluate the effect of the produced conduit on neural regeneration, a 10-mm sciatic nerve defect was created in rats, and experiments were carried out on six groups, i.e., sham control group (SC), negative control group (NC), nerve autograft group (NG), the biological conduit (AdMSCs + AM) group, the commercial PGA tube conduit (PGA) group, and the conduit only consisting of AM (AM) group. The effects of different nerve repair methods on the peripheral nerve and gastrocnemius muscle were evaluated by functional, histological, and immunohistochemical tests. When the number of myelinated axons was compared between the groups of AdMSCs + AM and PGA, it was higher in the AdMSCs + AM group (p < 0.05). The percentage of gastrocnemius collagen bundle area of AdMSCs + AM group was found to be statistically lower than the PGA group (p < 0.05). The muscle fiber diameter of AdMSCs + AM group was lower than that of the NG group, but significantly higher than that of the PGA group and the AM group (p < 0.001). Muscle weight index was significantly higher in the AdMSCs + AM group compared to the PGA group (p < 0.05). It was observed that nerve regeneration was faster in the AdMSCs + AM group, and there was an earlier improvement in pin-prick score and sciatic functional index compared to the PGA group and the AM group. In conclusion, the biological conduit prepared from the AdMSCs sheet and AM is regarded as a new biological conduit that can be used as an alternative treatment method to nerve autograft in clinical applications. [ABSTRACT FROM AUTHOR]

Amniotic membrane (AM) is an avascular structure composed of three different layers, which contain collagen, extracellular matrix, and biologically active cells (stem cells). Collagen, a naturally occurring matrix polymer, provides the structural matrix/strength of the amniotic membrane. Tissue remodeling is regulated by growth factors, cytokines, chemokines, and other regulatory molecules produced by endogenous cells within AM. Therefore, AM is considered an attractive skin-regenerating agent. This review discusses the application of AM in skin regeneration, including its preparation for application to the skin and its mechanisms of therapeutic healing in the skin. This review involved collecting research articles that have been published in several databases, including Google Scholar, PubMed, Science Direct, and Scopus. The search was conducted by using the keywords 'amniotic membrane skin', 'amniotic membrane wound healing', 'amniotic membrane burn', 'amniotic membrane urethral defects', 'amniotic membrane junctional epidermolysis bullosa', and 'amniotic membrane calciphylaxis'. Ultimately, 87 articles are discussed in this review. Overall, AM has various activities that help in the regeneration and repair of damaged skin. [ABSTRACT FROM AUTHOR]

Toxic epidermal necrolysis (TEN) is a devastating challenge faced by wound care practitioners. However surgical management with amniotic membrane graft (AMH) promises better results in scar healing, but there is a lack of additional intervention to reduce the high mortality rate in those patients who are admitted with poor prognosis clinical situation. There is supportive evidence that combination therapy with AMG and early high dose intravenous immunoglobulin (IVIG) as early management in TEN could improve the prognosis. In this study, the authors present a 61-year-old female who presented with a widespread macular lesion on the trunk, upper extremities and face. The patient was treated with early amniotic membrane grafting and high dose IVIG and achieved satisfactory results. Complete wound healing was achieved in 21 days. There were no acute or chronic complications, consisting of infection of the skin areas, gastrointestinal bleeding, pleural effusion, and severe ocular mucous membrane damage. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the effectiveness of cryopreserved human amniotic membrane transplantation (hAMT) in patients with infectious ulcerative keratitis. Methods: A retrospective cohort study based on medical records of patients who underwent hAMT procedure over a 2-year period (2020–2021) was conducted at Ege University Hospital, Turkey. Cryopreserved hAM was used. Best corrected visual acuity (BCVA) was measured with the Snellen chart. Detailed ophthalmological examination was performed at all visits. Treatment success was defined as complete healing including disappearance of corneal infiltrates, epithelial defect closure, reduction in anterior segment reaction. Results: A total of 13 eyes of 12 patients were diagnosed with ulcerative keratitis between 2020 and 2021. hAMT was performed in severe keratitis patients in which corneal infiltrates extending to middle to deep stromal and patients with neurotrophic component, in other words, having trouble for wound healing. The mean age was 60.9 ± 18.7 years, and 66.6% of the patients were male. Average ulcer horizontal diameter was 9.7 ± 2.8 mm. A single-layer human amniotic membrane (hAM) was transplanted 92.3% of eyes. Overlay technique was used in all patients for hAMT. The mean residence time of the amniotic membrane on the ocular surface was 14.1 ± 10.2 days. The mean time from the first presentation to the hAM transplantation was 9.7 ± 6.8 days. The average hospitalization time of all patients was 15 ± 12.1 days, pre-hAMT was 10.2 ± 7.3 days and post-hAMT was 4.8 ± 9.1 days. The mean follow-up time was 3 months. Recovery response was positive in 76.9%. Average healing time was 32.1 ± 14.04 days. Conclusions: hAMT was effective in the treatment of infectious ulcerative keratitis by accelerating and supporting wound healing. Performing hAM transplantation at the early stages of the disease and prolonged residence of membrane on the ocular surface increase the success of the treatment and moreover shortens hospitalization time. [ABSTRACT FROM AUTHOR]

Microbial control through alternative therapies, such as the amniotic membrane (AM) and antimicrobial photodynamic therapy (aPDT), has been gaining prominence with the advancement of bacterial resistance to conventional treatments. This study aimed to evaluate the antimicrobial effect of AM isolated and associated with aPDT using the PHTALOX® as a photosensitizer (PS) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms. The groups studied were: C+; L; AM; AM+L; AM+PHTX; and AM+aPDT. The irradiation parameters were 660 nm, 50 J.cm−2, and 30 mW.cm−2. Two independent microbiological experiments were carried out in triplicate, and the results were analyzed by CFU/mL counting and a metabolic activity test, both statistically analyzed (p < 0.05). The integrity of the AM was verified after the treatments by a scanning electron microscope (SEM). The groups AM, AM+PHTX, and, mainly, AM+aPDT showed a statistical difference when compared to C+ regarding the decrease in CFU/mL and metabolic activity. SEM analysis showed significant morphological alterations in the AM+PHTX and AM+aPDT groups. The treatments with AM isolated or associated with PHTALOX® were adequate. The association had potentiated the biofilm effect, and the morphological differences presented by AM after treatment did not hinder its antimicrobial effect, encouraging its use in biofilm formation locals. [ABSTRACT FROM AUTHOR]

Objective: To report the efficacy and long-term outcomes of treating the skin defects of aplasia cutis congenita (ACC) with cryopreserved amniotic membrane (AM). Method: Human amnion was obtained from the caesarean delivery of a full-term healthy pregnancy and processed in a sterile laminar flow hood, and cryopreserved in liquid nitrogen. The structure of the AM was investigated histologically and the viability of the epithelial cells was assessed after cryopreservation and compared with fresh AM and with AM preserved in phosphate-buffered saline (PBS) at 4°C. The cryopreserved AM was applied onto the lower limb skin defects of a one-month old baby with ACC. Timely AM changes were performed as necessary until the wounds healed. Results: The structure of the cryopreserved AM was intact, with little visible difference compared with fresh AM. The viability of the epithelial cells was partially lost but still much better retained than in those preserved in PBS at 4°C. The limb skin defects were gradually re-epithelialised upon application of the AM and were completely healed after one month. The 4-month and 2-year follow-ups presented good skin texture and colour, without hypertrophic scar formation. Conclusion: In this case study, cryopreservation of AM presented a well preserved stromal compartment and viable epithelial layer. It also offered features such as pain relief, good attachment and adhesiveness, improved wound healing and suppressed scar formation in the treatment of ACC skin defects. [ABSTRACT FROM AUTHOR]

The pathomechanisms of preeclampsia (PE), a complication of late pregnancy characterized by hypertension and proteinuria, and due to improper placentation, are not well known. Mesenchymal stem cells derived from the amniotic membrane (AMSCs) may play a role in PE pathogenesis as placental homeostasis regulators. PLACenta-specific protein 1 (PLAC1) is a transmembrane antigen involved in trophoblast proliferation that is found to be associated with cancer progression. We studied PLAC1 in human AMSCs obtained from control subjects (n = 4) and PE patients (n = 7), measuring the levels of mRNA expression (RT-PCR) and secreted protein (ELISA on conditioned medium). Lower levels of PLAC1 mRNA expression were observed in PE AMSCs as compared with Caco2 cells (positive controls), but not in non-PE AMSCs. PLAC1 antigen was detectable in conditioned medium obtained from PE AMSCs, whereas it was undetectable in that obtained from non-PE AMSCs. Our data suggest that abnormal shedding of PLAC1 from AMSC plasma membranes, likely by metalloproteinases, may contribute to trophoblast proliferation, supporting its role in the oncogenic theory of PE. [ABSTRACT FROM AUTHOR]

Objective: One of the main approaches to preventing skin ageing is to protect fibroblast cells from oxidative stress. The promoting effect of the human amniotic membrane extract (hAME) on re‐epithelization, proliferation and migration of cells in wound healing has been already well studied. This experimental study aimed to investigate the antioxidant activity of hAME against hydrogen peroxide (H2O2)‐induced dermal fibroblast damage. Methods: Here, to establish the ageing model, human foreskin fibroblasts (HFFs) were exposed to 200 μM H2O2 for 2 h. HFFs were treated with 0.1 mg/ml AME for 24 or 48 h before or/and after H2O2 exposure. A total of 48 h following the H2O2 treatment, we measured cell proliferation, viability, senescence‐associated β‐galactosidase (SA‐β‐Gal), antioxidants and preinflammatory cytokine (IL‐6) levels, as well as the expression of senescence‐associated genes (P53 and P21). Results: The obtained results indicated that under oxidative stress, AME significantly increased cellular viability and not only promoted the cell proliferation rate but also attenuated apoptotic induction condition (p < 0.001). AME also significantly reversed the SA‐β‐Gal levels induced by H2O2 (p < 0.001). Additionally, both pre‐ and post‐treatment regimen by AME down‐regulated the expression of senescence marker genes (p < 0.001). Moreover, AME declined different oxidative stress biomarkers such as superoxide dismutase and catalase and increased the glutathione amount. Conclusion: Altogether, our results indicated that AME had a remarkable antioxidant and antiageing activity as pre‐ and post‐treatment regimen, pointing to this compound as a potential natural‐based cosmeceutical agent to prevent and treat skin ageing conditions. [ABSTRACT FROM AUTHOR]

Type 1 diabetes mellitus (T1DM) is a complex metabolic disease characterized by a massive loss of insulin-producing cells due to an autoimmune reaction. Currently, daily subcutaneous administration of exogenous insulin is the only effective treatment. Therefore, in recent years considerable interest has been given to stem cell therapy and in particular to the use of three-dimensional (3D) cell cultures to better reproduce in vivo conditions. The goal of this study is to provide a reliable cellular model that could be investigated for regenerative medicine applications for the replacement of insulin-producing cells in T1DM. To pursue this aim we create a co-culture spheroid of amniotic epithelial cells (AECs) and Wharton's jelly mesenchymal stromal cells (WJ-MSCs) in a one-to-one ratio. The resulting co-culture spheroids were analyzed for viability, extracellular matrix production, and hypoxic state in both early- and long-term cultures. Our results suggest that co-culture spheroids are stable in long-term culture and are still viable with a consistent extracellular matrix production evaluated with immunofluorescence staining. These findings suggest that this co-culture may potentially be differentiated into endo-pancreatic cells for regenerative medicine applications in T1DM. [ABSTRACT FROM AUTHOR]

Human amniotic membranes (hAMs) are extraembryonic tissues currently employed in the treatment of many ocular and systemic diseases. Several reports indicate that hAMs can suppress the signaling pathway of tissue growth factor beta (TGF-β), a cytokine that plays a major role in the pathogenesis of proliferative vitreoretinopathy (PVR) through the induction of epithelial-mesenchymal transition (EMT) in exposed retinal pigmented epithelium (RPE) cells. The present study was conducted to evaluate the efficacy of a modified vitrectomy procedure (hAMP-V) involving the extensive coverage of exposed RPE with hAM patches to prevent postoperative PVR in a series of 15 cases of retinal detachment complicated by severe preoperatory PVR. The primary outcome was to assess the rate of successful retinal reattachment of a single hAMP-V procedure at 6 months from silicone oil removal. Secondary outcomes included the collection of intraoperative data concerning the quantity, size, and scope of hAM patches, and the assessment of postoperative improvements in mean LogMar BCVA at 3 and 6 months. Successful retinal reattachment was obtained in 14 out of 15 eyes (93.3%). Surgical failure due to major recurrence of PVR occurred in 1 out of 15 eyes (6.7%). Postoperative improvements in mean LogMar BCVA were statistically significant (p < 0.05, paired t-test). No intraoperative and postoperative adverse effects were reported. The study helped to refine the surgical technique while also offering cues for future improvements. [ABSTRACT FROM AUTHOR]

Neurotrophic keratitis is a complex condition with a variety of aetiologies involving damage to the sensory nerves that innervate the cornea, resulting in potentially devastating ocular surface disease. Aetiologies include diabetes, prior herpesvirus keratitis, stroke or compressive damage to the trigeminal nerve, chronic ocular surface disease or contact lens overuse, among others. Management is multidisciplinary; of course, the underlying condition must be addressed and treated. The ocular surface must be stabilized, and depending on the severity, treatment options include artificial tears and lubrication, protective barriers (scleral lens, tarsorrhaphy), and growth factors that promote healing (serum tears, amniotic membrane). Finally, treatment can target restoration of corneal sensory innervation (cenegermin, corneal neurotization). Early recognition and treatment are important factors in reducing vision loss and long-term complications from neurotrophic keratitis. [ABSTRACT FROM AUTHOR]

Treatment of large bone defects with supraphysiological doses of bone morphogenetic protein‐2 (BMP‐2) has been associated with complications including heterotopic ossification (HO), inflammation, and pain, presumably due to poor spatiotemporal control of BMP‐2. We have previously recapitulated extensive HO in our rat femoral segmental defect model by treatment with high‐dose BMP‐2 (30 μg). Using this model and BMP‐2 dose, our objective was to evaluate the utility of a clinically available human amniotic membrane (AM) around the defect space for guided bone regeneration and reduction of HO. We hypothesized that AM surrounding collagen sponge would attenuate heterotopic ossification compared with collagen sponge alone. In vitro, AM retained more BMP‐2 than a synthetic poly(ε‐caprolactone) membrane through 21 days. In vivo, as hypothesized, the collagen + AM resulted in significantly less heterotopic ossification and correspondingly, lower total bone volume (BV), compared with collagen sponge alone. Although bone formation within the defect was delayed with AM around the defect, by 12 weeks, defect BVs were equivalent. Torsional stiffness was significantly reduced with AM but was equivalent to that of intact bone. Collagen + AM resulted in the formation of dense fibrous tissue and mineralized tissue, while the collagen group contained primarily mineralized tissue surrounded by marrow‐like structures. Especially in conjunction with high doses of growth factor delivered via collagen sponge, these findings suggest AM may be effective as an overlay adjacent to bone healing sites to spatially direct bone regeneration and minimize heterotopic ossification. [ABSTRACT FROM AUTHOR]

Introduction: Spontaneous rupture of the membranes any time beyond the 28th week of pregnancy but before the onset of labour is called Premature Rupture of Membranes (PROM). There are many techniques for diagnosing PROM that is based on both clinical assessments and biological studies. Each of these procedures has benefits and drawbacks. Since foetal urine is the major source of amniotic fluid, the presence of vagina and a high creatinine level may aid in the diagnosis of PROM. Aim: To determine the cut-off levels of serum creatinine in predicting the PROM. Materials and Methods: A cross-sectional study was conducted at Dr. DY Patil Medical College, Hospital and Research Centre in Pune, Maharashtra, India from June 1 2020 to June 5 2021. Two equal groups of 140 each pregnant women with gestational ages ranging from 28-42 weeks were created: one with a history of vaginal leaking (the study group, P), and the other with gestationally matched no leakage (the control group, C). Patient's details such as age, gestational age, and obstetric history were noted. Each patient was examined and cervicovaginal secretions were collected. A 5 mL of saline was flushed in the posterior fornix of vagina. This fluid with vaginal wash was aspirated and creatinine concentration from vaginal fluid was determined by Jaffee synthetic chemical colorimetric method. The Student's t-test and the Chi-square test were used to analyse the data. Results: The study group's vaginal fluid creatinine level was substantially higher (p-value<0.001). The cut-off value of 0.6 mg/dL had highest sensitivity 96.1% and specificity 100%. The area under the Receiver Operating Characteristics (ROC) curve was 0.917. The Positive Predictive Value (PPV) and Negative Predictive Value (NPV) of serum creatinine at cut-off 0.6 mg/dL was 100% and 96.3% respectively. Conclusion: Vaginal fluid creatinine is simple and easily available, which makes it acceptable to the candidate for a diagnosis of PROM. [ABSTRACT FROM AUTHOR]

PURPOSE: To assess the extent of inferior fornix shortening in conjunctivochalasis (CCh) and to evaluate whether fornix deepening reconstruction can restore the fornix tear reservoir in patients with CCh. MATERIALS AND METHODS: This was a retrospective review of five patients (3 unilateral and 2 bilateral eyes, total 7 eyes) with CCh who underwent fornix deepening reconstruction with conjunctival recession and amniotic membrane transplantation. Postsurgical outcome measures included changes in fornix depth with correlation to basal tear volumes, symptoms, corneal staining, and conjunctival inflammation. RESULTS: For the three patients with unilateral surgery, both the fornix depth (8.3 ± 1.5 mm) and wetting length (9.3 ± 8.5 mm) of the operative eyes were less than the fellow eyes (10.3 ± 1.5 mm and 10.3 ± 8.5 mm, respectively). At 5.3 ± 2.7 months (range 1.7-8.7) postoperatively, the fornix depth increased significantly by 2.0 ± 1.1 mm (P = 0.02). Deepening of the fornix depth was accompanied by overwhelming symptomatic relief (91.5%) that could be subdivided into complete relief (87.5%) and partial relief (4%) of symptoms, with blurred vision being the most notably relieved symptom (P = 0.03). Furthermore, superficial punctate keratitis and conjunctival inflammation were significantly improved at follow-up (P = 0.008 and 0.05, respectively). CONCLUSION: Deepening of the fornix to restore the tear reservoir is an important surgical objective that may change the tear hydrodynamic state to provide a stable tear film and improve outcomes in CCh. [ABSTRACT FROM AUTHOR]

Background: The fetal amniotic membrane is a biological graft with unique qualities which all lead to wound protection, reducing discomfort, and achieving adequate epithelialization. Materials and Methods: In this animal study, the second and third premolars of the mandible of 4 dogs were extracted. After 4 weeks, 20 mm of mandibular premolar site area were resected on both sides. The created defects on both sides were filled with xenograft. On one side, an amniotic membrane was placed over the graft particles and the reflected flap was sutured. The amount of bone formation in the defects was measured after 4 weeks for two of the dogs and after 8 weeks for the other two, using a caliper. Three histopathological samples from both sides were taken. The collected data were subjected to statistical analysis (Wilcoxon signed-rank and paired sample t-test) using SPSS software at a significant P = 0.05. Results: In the test group, the quantity of bone was 56.81, whereas in the control group bone quantity was 37.38 with statistically significant differences (P = 0.025). In the amniotic membrane group, the inflammation intensity after the graft procedure was moderate (50%) in comparison to the control group where the inflammation was severe (62.5%) (P = 0.041). Conclusion: The amniotic membrane can induce positive osteoinduction effects and be helpful in repairmen of bone defects such as the natural periosteum. [ABSTRACT FROM AUTHOR]

The potential clinical applications of human amniotic membrane (hAM) and human amniotic epithelial cells (hAECs) in the field of regenerative medicine have been known in literature since long. However, it has yet to be elucidated whether hAM contains different anatomical regions with different plasticity and differentiation potential. Recently, for the first time, we highlighted many differences in terms of morphology, marker expression, and differentiation capabilities among four distinct anatomical regions of hAM, demonstrating peculiar functional features in hAEC populations. The aim of this study was to investigate in situ the ultrastructure of the four different regions of hAM by means of transmission electron microscopy (TEM) to deeply understand their peculiar characteristics and to investigate the presence and localization of secretory products because to our knowledge, there are no similar studies in the literature. The results of this study confirm our previous observations of hAM heterogeneity and highlight for the first time that hAM can produce extracellular vesicles (EVs) in a heterogeneous manner. These findings should be considered to increase efficiency of hAM applications within a therapeutic context. [ABSTRACT FROM AUTHOR]

In human body, biomaterials can be used to accelerate the wound healing process. Amniotic membrane-derived secretory leukocyte protease inhibitor (SLPI) is one of the biomaterials which able to accelerate the wound healing process by its several functions such as protease inhibition, leukocytes activity control, growth factor regulation such as transforming growth factor β (TGF-β), and its anti inflammatory, anti bacterial also anti retroviral properties. Objectives: To analyzed the SLPI effect on the TNF-α expression and the quantity of new blood vessel in the incisional wound healing. SLPI are produced from Essericea coli TOP10 as the cloning host, BL21 (DE3) strains as the expression host and pET30a plasmids were used for the expression system construction. The incision wound was made in dorsal skin area, then treated with SLPI dose 0.03 cc, 0.45 cc, 0.06 cc. After treatment, the wound covered with hypafix. The control group only covered with hypafix. In the next four days, the dorsal skin area is biopsied, then the quantity of TNF-α expression and blood vessels was analyzed with hematoxylin eosin and immunohistochemistry staining. The SLPI 0.045 cc showed the higher quantity of new blood vessel compared the SLPI 0.03 cc and control group (p

AIM: To evaluate the therapeutic effect of amniotic membrane (AM) for covering high myopic macular hole associated with retinal detachment following failed primary surgery. METHODS: Seventeen eyes of 17 patients whose axial length was more than 29 mm suffered from macular hole (MH) or MH associated with retinal detachment (RD), and had previously surgery of pars plana vitrectomy (PPV) with internal limiting membrane (ILM) peeling and silicone oil (SO) tamponade. Half a year after the surgery, optical coherence tomography (OCT) showed that MH did not heal in all 17 eyes and RD was still maintained in 13 eyes of these 17 eyes. We performed SO removal combined with AM covering on macular area and C3F8 tamponade, and phacoemulsification combined with intraocular lens implantation simultaneously cataract eyes. We followed up these patients for one year. RESULTS: In all 17 eyes, SO was removed successfully, MHs were healed and RDs were reattached. One eye (5.89%, 1/17) had AM shifted half a month after surgery and underwent a second surgery to adjust the position of the AM and supplement C3F8. After surgery, the visual acuity (VA) improved in 15 eyes (88.24%, 15/17), no change in two eyes (11.76%, 2/17). No serious complications occurred in all eyes. CONCLUSION: AM covering is helpful to rescue the previous failure surgery of high myopic MH.

Diabetic wound is one of the most common and costly complication in diabetic patients. Hence, numerous studies have been carried out to discover a suitable approach to enhance the process of wound healing. Biological hydrogels are commonly utilized for wound healing due to their suitable properties among different materials available. Herein we investigated whether human amniotic membrane hydrogel (hAMH) loaded with human placental mesenchymal stem cells (PlaMSCs)-derived exosomes could promote healing in diabetic rats. Sixty diabetic rats were randomly assigned into the control group, hAMH group, exosome group, and hAMH+Exosome group. According to the phases of wound healing, sampling was done on days 7, 14, and 21 for further assessments. Our findings showed a significant increase in wound contraction rate, new epidermal length, fibroblast and blood vessel count, collagen density, and the levels of antioxidative factors (GSH, SOD, and CAT) in the treatment groups compared to the control group, with more pronounced effects observed in the hAMH+Exosome group. Furthermore, the levels of bFGF and VEGF gene expression significantly increased in each treatment group when compared to the control group, with the highest levels observed in the hAMH+Exosome group. This occurred as the hAMH+Exosome group showed a greater decrease in neutrophil count, the expression of TNF-α and IL-1β genes, and the levels of an oxidative factor (MDA) compared to the other groups. In summary, the combination of hAMH and PlaMSCs-derived exosomes was determined to have a more significant effect on healing diabetic wounds. • The hAMH loaded with PlaMSCs -derived exosomes accelerate wound healing. • The hAMH loaded with PlaMSCs-derived exosomes regulate the inflammatory process. • The hAMH loaded with PlaMSCs-derived exosomes improve the histological characteristics. • The hAMH loaded with PlaMSCs-derived exosomes improve antioxidant statues. • The hAMH loaded with PlaMSCs-derived exosomes upregulated regenerative genes. [ABSTRACT FROM AUTHOR]

The heavy chain (HC)-hyaluronan (HA)/pentraxin 3 (HC-HA/PTX3) complex is formed by tumor necrosis factor-stimulated gene-6 (TSG-6) catalyzing the covalent (ester bond) transfer of HC1 from inter-α-trypsin inhibitor (IαI) to HA followed by tight binding of PTX3. The presence of such a complex has been found in human amniotic membrane (AM) and is considered to be a major matrix component responsible for its anti‑inflammatory and anti‑scarring properties to promote regenerative healing. Because the therapeutic potentials of AM and umbilical cord (UC) are similar, we herein evaluated whether human UC also contains HC-HA/PTX3. Immunostaining of UC cross-sections showed abundant PTX3, HC1, HA, TSG-6, and bikunin. Western blot analysis suggested the presence of HC1 complex bound via a NaOH-sensitive bond and tightly bound to PTX3 multimer in UC and AM extracts but not in chorion and placenta extracts. HC-HA/PTX3 was purified from UC extract by successive runs of density gradient ultracentrifugation and verified the presence of HC1 but not HC2 or HC3 based on western blot analysis. These results suggest the presence of HC-HA/PTX3 complex in UC is similar to AM. • Heavy chain (HC)-hyaluronan (HA)/pentraxin 3 (HC-HA/PTX3) present in amniotic membrane. • HC-HA/PTX3 has anti‑inflammatory and anti‑scarring properties. • HC-HA/PTX3 is also present in umbilical cord. [ABSTRACT FROM AUTHOR]

Although the human amniotic membrane (hAM) has been demonstrated to promote angiogenesis, its efficacy in healing ischemic wounds remains unknown. Therefore, the current study aimed to evaluate the potential of hAM as a dressing for treating ischemic wounds. The inferior abdominal wall arteries and veins of male rats were divided, and an ischemic wound was created on each side of the abdominal wall. Of the two ischemic wounds created, only one was covered with hAM, and its wound healing effect was determined by measuring the wound area. Angiogenesis was assessed by measuring microvessel density (MVD). On day 5, the mean wound area changed from 400 mm2 to 335.4 (260–450) mm2 in the hAM group and to 459 (306–570) mm2 in the control group (p = 0.0051). MVD was 19.0 (10.4–24.6) in the hAM group and 15.1 (10.6–20.8) in the control group (p = 0.0026). No significant differences in local pro- and anti-inflammatory cytokine levels were observed between the two groups. Histological examination revealed no rejection of the transplanted hAM. Therefore, the hAM may serve as a novel wound dressing that can promote angiogenesis and healing in ischemic wounds. [ABSTRACT FROM AUTHOR]

AIM:To investigate the clinical effect of modified free conjunctival flap transplantation with limbal stem cells in the treatment of pterygium and its impact on tear film function.METHODS: A total of 60 pterygium patients admitted to our hospital from March 2017 to March 2021 were randomly divided into the control group and the observation group, with 30 cases in each group. The control group was treated with amniotic membrane transplantation, and the observation group was treated with modified free conjunctival flap transplantation with limbal stem cells. The treatment period of both groups was 21d. And then the clinical efficacy, operation duration, corneal wound repair time, xerophthalmia score, tear film rupture time value and basic tear secretion test value before and 1, 3mo after treatment, and the occurrence of adverse reactions were compared between the two groups.RESULTS: The total effective rate of clinical treatment in the observation group was higher than that of the control group(χ2=5.963, P=0.015), the operation duration in the observation group was longer than that for the control group(t=-2.643, P

Some tissues of the body such as the amniotic membrane and cheap and available medicinal plants such as Arnebia euchroma can be effective in treating burn wounds, reducing its infection, and healing time. The aim of the present study was to investigate the antimicrobial effects of a combination of amniotic membrane, Arnebia Euchroma (AE) extract, sesame oil, and bee wax on the growth of gram-positive and gram-negative bacteria in second-degree burn wounds. After receiving amniotic membrane tissue from pregnant women, this tissue was mixed with sesame oil, bee wax, and Arnebia Euchroma extract and applied to the burn wounds of mice model infected with E. coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii. This combination was applied in case and control groups, and microbial counting was then performed. According to the findings of the present study, the number of pathogens in wound samples of mice treated with this combination was less than in control mice. The results of this study also showed that the use of a combination of amniotic membrane, AE plant extract, sesame oil, and bee wax had a therapeutic effect on second-degree burn wounds. The use of this compound after further research is recommended for the treatment of burn wounds.

Elizabeth Yeu,1,2 Omar Hashem,3 Hosam Sheha3– 5 1Virginia Eye Consultants, Norfolk, VA, USA; 2Eastern Virginia Medical School, Norfolk, VA, USA; 3Research Institute of Ophthalmology, Cairo, Egypt; 4Florida International University Herbert Wertheim College of Medicine, & Glaucoma Research Organization, Miami, FL, USA; 5Manhattan Eye, Ear and Throat Hospital, Hofstra Northwell School of Medicine, New York, NY, USACorrespondence: Hosam Sheha, Department of Ophthalmology, Manhattan Eye, Ear and Throat Hospital, 210 E 64 Street, New York, NY, 10065, USA, Tel +1 917-810-9555, Email hoss88@gmail.comPurpose: To assess the effectiveness of cryopreserved amniotic membrane (CAM) after debridement in treating epithelial basement membrane dystrophy (EBMD) prior to cataract surgery.Methods: This pilot study included 2 treatment groups: a prospective study group of 9 subjects with significant EBMD who received debridement followed by self-retained CAM, and a retrospective, control group of 10 consecutive subjects who received debridement followed by a bandage contact lens (BCL). Slit-lamp photography after fluorescein staining were used to monitor healing. Corneal topography and IOL calculation were compared at baseline and 1 month after the procedure. Refraction and ocular surface stability were also compared after cataract surgery.Results: Corneal reepithelialization after debridement occurred in 4.6 ± 0.8 days in the study group and 6.8 ± 0.6 days in the control (p < 0.05). Corneal topography showed changes in curvature from 43.5 ± 1.2D at baseline to 44.6 ± 1.2D at 1 month in the study group and from 45.0 ± 0.6D to 45.7 ± 0.8D in the control (p = 0.38). Average change in IOL calculation was 1.56 D in the study group, compared to 0.95 D in control (p = 0.29). Post-cataract refraction in both groups was within ± 0.5 Diopter of the anticipated, and corneal surface remained stable without EBMD recurrence.Conclusion: Management of ocular surface disorders prior to cataract surgery stabilizes IOL calculation and reduces postoperative refractive surprises. CAM relatively accelerated healing after debridement; however, it was not better than BCL in stabilizing the ocular surface and improving visual outcome. The use of CAM in cases of EBMD remains speculative.Keywords: amniotic membrane, cataract, EBMD, ocular surface

Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), has become in the spotlight regarding the serious early and late complications, including acute respiratory distress syndrome (ARDS), systemic inflammation, multi-organ failure and death. Although many preventive and therapeutic approaches have been suggested for ameliorating complications of COVID-19, emerging new resistant viral variants has called the efficacy of current therapeutic approaches into question. Besides, recent reports on the late and chronic complications of COVID-19, including organ fibrosis, emphasize a need for a multi-aspect therapeutic method that could control various COVID-19 consequences. Human amniotic epithelial cells (hAECs), a group of placenta-derived amniotic membrane resident stem cells, possess considerable therapeutic features that bring them up as a proposed therapeutic option for COVID-19. These cells display immunomodulatory effects in different organs that could reduce the adverse consequences of immune system hyper-reaction against SARS-CoV-2. Besides, hAECs would participate in alveolar fluid clearance, renin–angiotensin–aldosterone system regulation, and regeneration of damaged organs. hAECs could also prevent thrombotic events, which is a serious complication of COVID-19. This review focuses on the proposed early and late therapeutic mechanisms of hAECs and their exosomes to the injured organs. It also discusses the possible application of preconditioned and genetically modified hAECs as well as their promising role as a drug delivery system in COVID-19. Moreover, the recent advances in the pre-clinical and clinical application of hAECs and their exosomes as an optimistic therapeutic hope in COVID-19 have been reviewed. Graphical abstract

Purpose: To formulate a treatment algorithm for the management of descemetocele. Methods: This was a prospective interventional study that was conducted at a tertiary eye-care center. All consecutive cases of descemetocele during the study period (April 1, 2017–March 31, 2018) were evaluated for the following parameters: age, sex, previous medical or surgical therapy, risk factors, preexisting ocular diseases, location, site and size of descemetocele, interventions undertaken, visual acuity, and the fellow eye status. The surgical modalities and fellow eye status were correlated individually with therapeutic and functional outcomes, based on which a treatment algorithm was formulated. Results: The study included 24 eyes of 24 patients (19M, 5F) with a median age of presentation of 45 years. The mean follow-up duration was 6.79 ± 3.97 months (3–12 months). The most common cause of descemetocele was microbial keratitis (66.66%), and most cases were central (50%), small (58.33%), and non-perforated (79.16%). The surgical interventions undertaken were cyanoacrylate glue (CG, 37.5%), penetrating keratoplasty (PKP, 33.33%), patch graft (16.66%), and deep anterior lamellar keratoplasty (DALK, 12.5%). Therapeutic success was noted in 13/24 eyes (54.16%). Final visual acuity > 3/60 was seen in 25% cases. Suboptimal therapeutic (P = 0.07) and visual (P = 0.34) outcomes were noted in subjects with non-functional fellow eye. Conclusion: PKP was preferred for descemetoceles with active microbial keratitis and extensive infiltrates, while CG and DALK were undertaken for healed microbial keratitis, neurotrophic keratitis, and ocular surface disorders with partial limbal stem cell deficiency (LSCD). For total LSCD, amniotic membrane graft was preferred.

Context: The use of human-derived amniotic barrier membranes in regenerative procedures along with allograft like demineralized freeze-dried bone allograft (DFDBA) has displayed promising outcomes. Aims: The objective of the current research was to clinically and radiographically evaluate and compare amniotic membrane (AM) and collagen membrane (CM) in combination with open flap debridement (OFD) along with DFDBA in the management of human periodontal intrabony defects. Settings and Design: The present research-based study was a randomized controlled clinical trial of 6 months duration. Materials and Methods: A total of twenty chronic periodontitis patients with intrabony defects were randomly divided into two groups: Group 1 treated with OFD + DFDBA + AM and Group 2 with OFD + DFDBA + CM. Clinical parameters such as plaque index (PI), gingival index (GI), pocket probing depth (PPD), and relative attachment level were recorded and evaluated at baseline, 3 months, and 6 months. The relative intrabony defect component level (rIBD) was recorded at baseline and after 6 months employing a millimeter X-ray grid. Statistical Analysis Used: For comparison of clinical parameters and radiographic parameters at various intervals unpaired t-test, repeated measures ANOVA test, and paired t-test were used. The P value was taken statistically significant when less than 0.05 (P < 0.05). Results: A statistically significant difference in PI, GI, PPD, and RAL for both the groups were found on intragroup comparison between baseline, 3 months, and 6 months (P < 0.05). On intragroup comparison, the difference in mean rIBD was statistically significant from baseline to 6 months. However, for all the clinical and radiographic parameters, no statistically significant difference was noted between both the groups (P > 0.05). Conclusions: In the treatment of intrabony defects AM being an allograft can be considered an alternative novel membrane for regenerative procedures.

PURPOSE: To describe the ocular manifestations in the acute stage of Stevens-Johnson syndrome/ toxic epidermal necrolysis. MATERIALS AND METHODS: We reviewed all the medical records of patients diagnosed with Stevens-Johnson syndrome/ toxic epidermal necrolysis and erythema multiforme between 2012 and 2019. Demographics, ophthalmic manifestations, and the treatment given systemically and locally were reviewed and analyzed. RESULTS: We had forty-five patients admitted to our hospital between Jan 2012 to Dec 2019 with SJS/TEN as a diagnosis.Twenty-six (57.5%) of them were females, and 19(42.2%) were males. The mean age was 27.5 years.Forty (88.9%) of our cases were diagnosed as Stevens-Johnson syndrome, and five (11.9%) as toxic epidermal necrolysis.We found antiepileptics as a triggering agent in thirteen cases (28.8%).Fever (84.4%) and mucosal lesions (86.7%) were the most common presenting symptom.We found ocular symptoms in only 22 (48.9%) patients.The treating physicians referred only thirty-one cases to the ophthalmologist, out of which 22 cases were referred within three days of admission.The most common ocular involvement was conjunctival congestion (69%).Ocular grading showed that mild grade included 42.9%, moderate grade 28.6%, and severe grade 28.6% of the cases.The ocular treatment involved medical management with lubricating drops (100%), topical steroids (58.6%), and topical antibiotics (68.9%). Five individuals with a severe grade of ocular involvement underwent amniotic membrane transplantation. CONCLUSION: Ocular examination and grading are essential in the acute stage of SJS/TEN. It helps the ophthalmologist recognize the sentinel findings and institute appropriate treatment in the acute stage as early as possible.

Objective: To report a case of bilateral Mooren’s ulcer with impending corneal perforation in the right eye and perforated peripheral corneal ulcer in the left eye which successfully underwent Tenon’s patch graft (TPG) with multilayered amniotic membrane grafting (AMG) as globe salvaging procedure during COVID-19 pandemic. Methods: Conjunctival resection was done on both the eyes followed by autologous Tenon’s grafting with overlay amniotic membrane grafting in the left eye with perforation and multilayered AMG with AMG overlay in the right eye with impending perforation. Post-operatively topical antibiotics and steroids were prescribed. Results: At one month follow-up, the Tenon’s graft and multilayered AMG were well-integrated with the surrounding healthy cornea with a good anatomical and visual outcome. Conclusion: Autologous Tenon’s patch graft is a simple and cost-effective alternative to preserve globe integrity and prevent complications in emergency cases when immediate access to cornea transplantation is practically challenging.

Purpose: To describe the microscopic epithelial changes and the clinical outcomes of a patient treated with amniotic membrane eye drops (AMED) because of a persistent epithelial defect (PED) and a partial limbal stem cell deficiency (LSCD) after simple limbal epithelial transplantation (SLET) and deep anterior lamellar keratoplasty (DALK). Observations: A 72-year-old patient, who had previously undergone SLET and DALK due to a total LSCD, presented with a PED related to a partial LSCD, and was treated with AMED for one month. We evaluated the patient's visual acuity, the Oxford grading scale, the Wong-Baker Pain Rating Scale, and in vivo confocal microscopy, both at baseline and 3 months after the end of treatment. Visual acuity improved from 0.5 to 0.4 LogMAR, the Oxford grading scale changed from grade III to grade I and the Wong-Baker Pain Rating Scale from grade 4 to grade 1. The corneal surface, which initially showed conjunctival characteristics over approximately 50% of the whole area, consisted mainly (75%) of mature corneal epithelium 3 months after the end of treatment. Conclusions and importance: While improving symptoms and clinical characteristics, AMED was also able to restore the normal corneal epithelium's morphology in a case of partial LSCD after SLET and DALK.

Introduction: Spontaneous rupture of the membranes any time beyond the 28th week of pregnancy but before the onset of labour is called Premature Rupture of Membranes (PROM). There are many techniques for diagnosing PROM that is based on both clinical assessments and biological studies. Each of these procedures has benefits and drawbacks. Since foetal urine is the major source of amniotic fluid, the presence of vagina and a high creatinine level may aid in the diagnosis of PROM. Aim: To determine the cut-off levels of serum creatinine in predicting the PROM. Materials and Methods: A cross-sectional study was conducted at Dr. DY Patil Medical College, Hospital and Research Centre in Pune, Maharashtra, India from June 1 2020 to June 5 2021. Two equal groups of 140 each pregnant women with gestational ages ranging from 28-42 weeks were created: one with a history of vaginal leaking (the study group, P), and the other with gestationally matched no leakage (the control group, C). Patient’s details such as age, gestational age, and obstetric history were noted. Each patient was examined and cervicovaginal secretions were collected. A 5 mL of saline was flushed in the posterior fornix of vagina. This fluid with vaginal wash was aspirated and creatinine concentration from vaginal fluid was determined by Jaffee synthetic chemical colorimetric method. The Student’s t-test and the Chi-square test were used to analyse the data. Results: The study group’s vaginal fluid creatinine level was substantially higher (p-value

The amniotic membrane (AM) is the inner part of the placenta. It has been used therapeutically for the last century. The biological proprieties of AM include immunomodulatory, anti-scarring, anti-microbial, pro or anti-angiogenic (surface dependent), and tissue growth promotion. Because of these, AM is a functional tissue for the treatment of different pathologies. The AM is today part of the treatment for various conditions such as wounds, ulcers, burns, adhesions, and skin injury, among others, with surgical resolution. This review focuses on the current surgical areas, including gynecology, plastic surgery, gastrointestinal, traumatology, neurosurgery, and ophthalmology, among others, that use AM as a therapeutic option to increase the success rate of surgical procedures. Currently there are articles describing the mechanisms of action of AM, some therapeutic implications and the use in surgeries of specific surgical areas, this prevents knowing the therapeutic response of AM when used in surgeries of different organs or tissues. Therefore, we described the use of AM in various surgical specialties along with the mechanisms of action, helping to improve the understanding of the therapeutic targets and achieving an adequate perspective of the surgical utility of AM with a particular emphasis on regenerative medicine.

Synopsis: Using a dehydrated human amniotic membrane along with an amniotic suspension allograft, an effective therapy for complex rectovaginal fistula is demonstrated. [ABSTRACT FROM AUTHOR]

Background: Burn injuries are associated with significant mortality and morbidity around the world. The care of burn wounds requires a great amount of medical resources, therefore it is important to find a wound dressing that accelerates wound healing and is readily available. Objectives: This review article will provide an overview of the literature and background of the usage of amniotic membranes in burn patients. We will also discuss its properties of preventing infections, relieving pain, how it promotes and accelerates wound healing. Discussion: Human amniotic membrane is now used around the world as a relatively cheap temporary dressing for burn wounds. It has been shown to promote wound healing, create a moist environment, reduce pain intensity, reduce scar formation, prevent water and electrolyte disturbances and also reduce the risk of infections. Research has also shown that it contains antimicrobial properties that could be of great benefit in burn patients and is compatible to use in developing countries because it is readily available, easy to obtain and sterilize, able to cover wounds of large size, protects the wound from excessive water and electrolyte loss, reduces pain intensity, requires fewer dressing changes and is also more cost effective than conventional dressings. Conclusion: Amniotic membrane is a biological dressing that can be useful in the treatment of burn wounds. Further research should be conducted to investigate and understand the mechanisms of amniotic membrane for burn and wound care. [ABSTRACT FROM AUTHOR]

Human term placenta and other postpartum-derived biological tissues are promising sources of perinatal cells with unique stem cell properties. Among the massive current research on stem cells, one medical focus on easily available stem cells is to exploit them in the design of immunotherapy protocols, in particular for the treatment of chronic non-curable human diseases. Type 1 diabetes is characterized by autoimmune destruction of pancreatic beta cells and perinatal cells can be harnessed both to generate insulin-producing cells for beta cell replenishment and to regulate autoimmune mechanisms via immunomodulation capacity. In this study, the strong points of cells derived from amniotic epithelial cells and from umbilical cord matrix are outlined and their potential for supporting cell therapy development. From a basic research and expert stem cell point of view, the aim of this review is to summarize information regarding the regenerative medicine field, as well as describe the state of the art on possible cell therapy approaches for diabetes. [ABSTRACT FROM AUTHOR]

Producing tremendous amounts of stress and financial burden on the global patient population and healthcare systems around the world, most current modalities of treatment for musculoskeletal ailments often do not address the etiopathogenetic causes of these disorders. Regenerative medicine for musculoskeletal disorders relies on orthobiologics derived from either allogenic or autologous sources. Multiple drawbacks are associated with autologous sources, including donor-site morbidity, a dearth of studies, and variability in both patient reported and clinical/functional outcomes. On the other hand, allogenic sources address several of these concerns, and continue to be a suitable source of mesenchymal stem cells (MSCs). This review qualitatively reports both the preclinical and clinical outcomes of publications studying the applications of umbilical cord (-derived Wharton's jelly), amniotic suspension allograft, amniotic membrane, and amniotic fluid in musculoskeletal medicine. A systematic review was conducted utilizing the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines on studies published between January 2010 and October 2022 that used allogeneic perinatal tissues. Further randomized controlled clinical studies are necessary to properly evaluate the safety and efficacy of these tissues in orthopedic surgery. [ABSTRACT FROM AUTHOR]

Covid pandemic affected donation activities worldwide, especially for living donation due to the lack of elective surgery. Moreover, the number of heart-beating and non-heart beating donors has recorded a decrease. Fondazione Banca dei Tessuti di Treviso (FBTV) is a non-profit healthcare organisation, located in Veneto Region, tasked with procurement, processing, preserving, validating and distributing human tissue for clinical use. During Covid-19 outbreak, operations in FBTV have never stopped and a great effort was required to maintain a standard trend of activity. The aim of this study was to describe the impact of Sars-CoV-2 on the activity of a multitissue bank in Italy. Moreover, we investigated the presence of the virus in tissues retrieved from two Sars-CoV-2 positive cadaver donors. Our survey demonstrated that the transplantation network of Veneto Region has positively reacted to the pandemic scenario, thanks to the effort of all personnel involved. Statistical analyses underlined that most of the activities of the tissue bank were unaffected during the Sars-CoV-2 pandemic. [ABSTRACT FROM AUTHOR]

Purpose: To compare the effects on adhesive and structural properties of newer preservation conditions to those obtained with an established, standardized protocol (dimethyl sulfoxide at −180 °C). In attempt to simplify and enhance the safety of the procedure, we tested dextran-based freezing medium and a dry condition (no medium) at temperatures of −80 °C. Methods: Five patches of human amniotic membrane were obtained from three different donors. For each donor, five preservation condition were tested: dimethyl sulfoxide at −180 °C, dimethyl sulfoxide at −80 °C, dextran-based medium at −180 °C, dextran-based medium at −80 °C and dry freezing at −80 °C (no medium). At the end of four months storage period, adhesive properties and structure were analyzed. Results: None of the newer preservation protocols showed differences in adhesive and structural properties of the tissues. The stromal layer always kept its adhesiveness, while both structure and basement membrane were not altered by any the preservation protocol. Conclusions: Switching from liquid nitrogen cryopreservation to −80 °C would reduce manipulation, simplify the procedure, making it also cheaper. The use of dextran-based freezing medium or no medium at all (dry condition) would avoid the potential toxicity of the dimethyl sulfoxide-based freezing media. [ABSTRACT FROM AUTHOR]

A perfect graft for wound care must be readily available without affecting the immune response, covering and protecting the wound bed. Considering previous studies have already established the use of hyaluronic acid (HA) for the treatment of wounds but the data presented on the amniotic membrane (AM) and its promising effects on healing still requires further investigation, this study aimed to evaluate the effects of the application of a decellularized amniotic membrane solubilized with hyaluronic acid on the healing process of cutaneous wounds on the 7th and 14th day, to evaluate the evolution of the wound and the inflammatory phases in these two times. Cutaneous lesions were excised from the dorsal region and 96 Wistar rats were divided into four groups: I–Excisional wound (EW); II–EW + AM; III–EW + HA; IV–EW + AM + HA. The present study demonstrated that the proposed combined therapy favors the tissue repair process of the epithelial lesion. Results showed a reduction in pro-inflammatory cytokines, an increase in anti-inflammatory cytokines, an increase in TGF-β, and attenuation of oxidative stress, reducing the acute inflammatory response and promoting the beginning of tissue repair. We concluded that the proposed therapies accelerated the inflammatory process with anticipation of the repair phase. [ABSTRACT FROM AUTHOR]

Objectives: The objective of this study was to evaluate the efficacy of cell therapy using human amniotic epithelial stem cells (hAESCs) for the treatment of premature rupture of membranes (PROM) in vitro. Design: Using the amniotic pore culture technique (APCT), we mimicked the environment of PROM in vitro, thus enabling the observation of the healing process of hAESC-treated amniotic membranes. Materials: Amniotic membrane samples were collected from placentas of pregnant women who underwent elective cesarean sections. APCT model and isolated hAESCs were used in this study. All patients who participated in this study provided their written informed consent prior to the commencement of the study. Settings: To create the APCT model in vitro, isolated amniotic membranes were punched to create 5 mm diameter circles and re-punched to form a 1-mm pore at the center. Membranes were cultured in α-minimal essential medium, and the hAESCs were collected and cultured as well. Subsequently, the APCT models were divided into two groups: hAESC treated and control. Methods: Within the culture period, pore sizes were calculated to evaluate the degree of tissue regeneration in both groups. We then evaluated the histology, cell density, and epithelial thickness of the regenerated tissues. Statistical analyses were performed using SPSS software ver. 20.0 (IBM, Armonk, NY, USA) with repeated-measures one-way analysis of variance or paired samples t test. The significance level was set at p < 0.05. Results: As per the evaluation of the APCT model in vitro, the pore size in the hAESC-treated group reduced by 62.2% on day 6 (62.2 ± 0.19, n = 24), whereas in the control group, it shrank by only 36.8% (p < 0.05) (36.8 ± 0.19, n = 24). Furthermore, the epithelial thickness in the amniotic epithelial stem cell-treated group (10.08 ± 1.26 μm, n = 8) was significantly higher than that in the control group (5.87 ± 0.94 μm, n = 8). Cell density in the regenerated tissue in the amniotic epithelial stem cell-treated group (57 ± 2.77, n = 8) was significantly higher than that in the control group (49 ± 2.23, n = 8). Limitations: In this study, we did not explore the molecular mechanisms by which hAESCs participate in membrane healing in the APCT model. Although our results showed a significant difference, this difference was not too obvious. Therefore, further research on the mechanisms of hAESCs is needed, with more amniotic tissues and APCT samples being tested. Conclusions: We developed an APCT model to investigate the PROM conditions in vitro. By implanting donor hAESCs in the pores of the APCT model, we observed that hAESCs seeding accelerated pore healing in vitro. Thus, hAESCs may be a valuable source of cells for cell therapies in regenerative medicine. [ABSTRACT FROM AUTHOR]

Background/aim: The goal of the present study was to investigate the effects of an acellular amniotic membrane loaded with autologous platelet-rich plasma on bone healing biochemically by measuring serum factors including calcium and alkalinity in a dog model. Materials and methods: Fifteen dogs were used in the current study. The animals were divided into three groups of five dogs each. A transverse osteotomy was induced in the midshaft of the left tibia of all animals. The first group was treated with acellular amniotic membrane only by wrapping it around the fracture site. The second group was given acellular amniotic membrane and platelet-rich plasma to treat them, while the third group (the control) was left alone and not treated Whole blood was drawn from all groups on the 0th day prior to surgery as well as on the 14th, 28th, 42nd, and 56th days following surgery. Physiological data was then reported. in order to estimate calcium and alkaline phosphatase at various phases of fracture healing. Results: the results of the present study illustrated that the serum alkaline phosphate and calcium levels increased (p=0.01) on days 14 and 28, followed by a decreasing trend after day 28 to day 56. And their levels were higher in group AM+PRP than in other groups (39.4 ± 1.21, 15.79±0.66 respectively). When compared to the control and AM groups, the rate of formation of new bone tissue in Group AM+PRP was the fastest. It was concluded that serum alkaline phosphatase and calcium were found to be helpful biochemical indicators for evaluating bone development and contributed to proving the superiority of using the amniotic membrane loaded with platelet-rich plasma in promoting bone healing. These biochemical parameters, in conjunction with clinical and radiographic examinations, provide reliable information about the degree of bone healing. [ABSTRACT FROM AUTHOR]

Objectives: The objective of the study was to evaluate the technique of tarsorrhaphy combined with sutureless amniotic membrane transplantation in the treatment of resistant ocular surface pathologies. Methods: In this study, the post-operative results of patients who underwent tarsorrhaphy combined with sutureless amniotic membrane between May 2016 and July 2021 were evaluated. In this technique, we have placed the amniotic membrane on the ocular surface, and then simple tarsorrhaphy has been performed. We used this technique in our six patients during COVID-19 pandemic period at the bedside. The outcomes of corneal epithelial healing, visual acuity, pain score, patient tolerance, and adverse events were evaluated. Results: Fourteen patients (eight females and six males) were included in the study. The mean age of the patients was 65.7 years (range 9-96 years). The mean follow-up period was 9 months (range 6-12 months). In all cases, epithelial defect healed completely and it was seen that this new technique facilitated resolution of pain and inflammation and promoted epithelialization in our 14 cases with resistant ocular surface pathologies. Post-operative pain score was significantly lower than pre-operative pain score (p<0.05). Conclusion: Our results demonstrate that tarsorrhaphy with sutureless amniotic membrane accelerates the ocular surface healing process and increases the efficiency of amniotic membrane. This method has an important advantage that it can be performed at bedside. Patients and ophthalmologists need such an innovative treatment which is effortless, effective, and practical especially for the pandemic period. [ABSTRACT FROM AUTHOR]

Objectives/Hypothesis: Vocal fold (VF) scar and sulcus cause severe vocal problems, but optimal methods have not been established. Total replacement of the mucosa is required particularly for cases in which the whole lamina propria is occupied by severe fibrosis and vibratory function is totally lost. The amniotic membrane (AM) has been proven to have regenerative potential, as it contains stem cells and growth factors. The current study investigated the biocompatibility and effects of AM for regeneration of the VF mucosa. Study Design: In vitro and in vivo studies. Methods: Vocal fold fibroblasts (VFFs) from 13 Sprague–Dawley rats were seeded on AM and subjected to histology and immunohistochemistry, and gene expressions in the VFFs on AM were examined in in vitro study. Twelve New Zealand White rabbits were used in in vivo study. VFs were stripped down and were reconstructed with AM. The regenerative effects were examined 3 months later by histological examination. Results: In vitro study indicated VFFs survived on AM and stained positively for Ki67, vimentin, and fibronectin. Gene expressions of Has1, Has2, and Hgf were significantly increased in the VFFs on AM compared with the other groups. The in vivo study indicated AM‐transplanted VFs showed a significantly higher density of hyaluronic acid and lower density of collagen compared with sham VFs. Conclusions: The current preliminary study suggests biocompatibility and possible regenerative effects of AM for VFs. Level of Evidence: NA Laryngoscope, 132:2017–2025, 2022 [ABSTRACT FROM AUTHOR]

We questioned the relevance of evaluating residual cell viability in human amniotic membrane (hAM) after its cryopreservation since cell survival is controversial and its ability to act as a matrix (including the presence of growth factors and cytokines) appears to be most important for tissue regeneration purposes. We also discussed the usefulness of osteodifferentiating amniotic cells in whole hAM for bone repair applications. We have evidence that determining residual cell viability after cryopreservation and hAM osteodifferentiation is not justified. [ABSTRACT FROM AUTHOR]