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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.

Background: The use of human amniotic membrane transplantation is rapidly increasing for the management of various ocular surface disorders. Despite its numerous advantages, amniotic membrane is not widely available due to the lack of awareness among ophthalmologists regarding its preparation and preservation techniques. Purpose: To provide an instructional video demonstrating the technique of harvesting, preparation, preservation of human amniotic membrane, and its uses in the management of ocular surface diseases. Synopsis: The amniotic membrane is the innermost layer of the human placenta. Properties of the amniotic membrane, like the presence of various growth factors, anti-inflammatory and anti-angiogenic factors, and its low immunogenicity, contribute to its ability to promote epithelial growth and differentiation with the reduction in fibrosis during healing. To harvest amniotic membrane, informed consent is obtained from a patient undergoing an elective caesarian section, and the donor is screened to exclude the risk of infections. Under sterile precautions, the amniotic membrane is separated from the chorion and washed free of blood clots. With the epithelial surface up, the amniotic membrane is spread uniformly without folds or tears on individually sterilized 0.22 μm nitrocellulose filter papers of the required sizes. The prepared filter paper with the adherent amniotic membrane is placed in freshly prepared Dulbecco's Modified Eagle's Medium and stored at −80°C. The amniotic membrane can be used for surgical procedures like symblepharon release, pterygium or dermoid excision, perforated ulcers, nonhealing epithelial defects, etc. Highlights: Instructional videos demonstrating the preparation and preservation of amniotic membrane are very sparse. This video clearly explains how any ophthalmologist can learn to prepare and preserve the human amniotic membrane and gives a glimpse of its properties, advantages, and scope of use. Video Link: https://youtu.be/8HfSjXUKoiY

A complex extracted from the amniotic membrane in humans reduces post-surgical pain in mice by directly inhibiting pain-sensing neurons., Competing Interests: JZ, JC No competing interests declared, (© 2024, Zhang and Cheng.)

Introduction: Localized prostate cancer (PCa) is one of the most common malignancies in the United States. Despite continued refinement of robot assisted radical prostatectomy (RARP) surgical methods, post-surgical erectile dysfunction and urinary incontinence remain significant challenges due to iatrogenic injury of local nervous tissue. Thus, the development of therapeutic strategies, including the use of biologic adjuncts to protect and/or enhance recovery and function of nerves following RARP is of growing interest. Perinatal tissue allografts have been investigated as one such biologic adjunct to nerve sparing RARP. However, knowledge regarding their clinical efficacy in hastening return of potency and continence as well as the potential underpinning biological mechanisms involved remains understudied. Thus, the objective of this literature review was to summarize published basic science and clinical studies supporting and evaluating the use of perinatal allografts for nerve repair and their clinical efficacy as adjuncts to RARP, respectively., Methods: The literature as of May 2024 was reviewed non-systematically using PubMed, EMBASE, Scopus, and Web of Science databases. The search terms utilized were "robotic prostatectomy", "prostate cancer", "nerve sparing", "perinatal tissue", "allograft", "potency", and "continence" alone or in combination. All articles were reviewed and judged for scientific merit by authors RP and JM, only peer-reviewed studies were considered., Results: Eight studies of perinatal tissue allograph use in RARP were deemed worthy of inclusion in this nonsystematic review., Conclusions: Incontinence and impotence remain significant comorbidities despite continued advancement in surgical technique. However, basic science research has demonstrated potential neurotrophic, anti-fibrotic, and anti-inflammatory properties of perinatal tissue allografts, and clinical studies have shown that patients who receive an intra-operative prostatic perinatal membrane wrap have faster return to potency and continence., (© 2024. The Author(s).)

Toxic epidermal necrolysis (TEN) is a severe, often drug-induced mucocutaneous reaction characterized by widespread epidermal necrosis and detachment. Ocular involvement is common and can lead to significant morbidity if not addressed promptly. This case report presents a 10-year-old female with fever, rash, and severe ocular symptoms. Early use of Prokera, an amniotic membrane device, was instrumental in managing her ocular condition. After discharge and follow-up care, the patient showed marked improvement. This case highlights the importance of early recognition and intervention in ocular manifestations of TEN to prevent long-term complications., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. RMS-MH/IRB/ 2024-846 issued approval 2024-846. This research study, "Early Recognition and Management of Ocular Manifestations of Toxic Epidermal Necrolysis in a Pediatric Patient: A Case Report," has been reviewed and approved by the Institutional Review Board (IRB) under protocol number RMS-MH/IRB/2024-846, in compliance with ethical standards for research involving human subjects. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Abulfateh et al.)

Chronic wounds result from the body's inability to heal, causing pain, pathogen entry, limited treatment options, and societal burden. Diabetic foot ulcers are particularly challenging, often leading to severe complications like leg amputation. A clinical study tested AMNIODERM+ ® , a new device with a lyophilized human amniotic membrane (HAM), on chronic diabetic foot ulcers. Participants had diabetic neuropathic or neuroischemic leg wounds (2-16 cm 2 ) unhealed by 20% after six weeks of standard care. This study showed significant wound healing improvements with AMNIODERM+ ® . The median wound size reduction after 12 weeks was 95.5%, far exceeding the null hypothesis of 20% change. Additionally, 65% of patients achieved complete ulceration healing, surpassing the 50% efficacy requirement. The median time to full closure was 11.4 weeks, with the proportion of completely healed patients rising progressively, reaching 55% by week 11. These findings, from the clinical trial "Freeze-dried amniotic membrane in the treatment of nonhealing wounds", suggest AMNIODERM+ ® as a promising future treatment for chronic diabetic foot ulcers. The published results were obtained as part of a clinical trial entitled "Freeze-dried amniotic membrane in the treatment of nonhealing wounds: a single-arm, retrospectively-perspective clinical trial", EUDAMED Nr. CIV-SK-22-10-041146.

Aim: To investigate the effect of adding autologous serum eye drops to the postoperative regime after amniotic membrane transplantation for severe persistent corneal ulcers., Methods: Forty eyes of 40 patients with persistent corneal ulcers were randomly assigned to artificial tears (sodium hyaluronate 0.2%, ATs group, n =20) or autologous serum eye drops (ASEDs, n =20) following treatment with amniotic membrane transplantation. Digital slit lamp images were acquired from all patients before and 30d post treatment. The area with fibrovascular tissue was calculated using Image J. Central corneal sensitivity was assessed by Cochet-Bonnet aesthesiometry before and one month after treatment. Scar tissue transparency was assessed with a novel optical densitometry., Results: Mean age of patients was 61.65±16.47y and 57.3±19.11y in the ATs group and ASEDs group, respectively. Twenty-two male and 18 female patients were included in the study. The improvement in visual acuity was significantly greater in the ASEDs group (0.14±0.04) than the ATs (0.08±0.04; P =0.00046). Cochet-Bonnet aesthesiometry improved significantly after treatment with a similar rate between groups. There were no statistically significant differences in the area of postoperative fibrovascular tissue between the two groups ( P =0.082). The success rate in the two groups was similar. The difference in densitometry between the ATs and ASEDs group was statistically significant ( P =0.042) with greater reduction from baseline in the ASEDS group., Conclusion: Autologous serum eye drops can lead to better visual acuity, more stable results and improved densitometry and should be considered in the postoperative care following amniotic membrane transplantation., Competing Interests: Conflicts of Interest: Hristova R, None; Yankova P, None; Markov G, None; Oscar A, None; Zdravkov Y, None., (International Journal of Ophthalmology Press.)

Objective: To assess the feasibility of using a novel device designed for minimally invasive suturing to anchor fetal membranes to the uterine wall and to close surgical defects after fetoscopy., Methods: We tested the WestStitch™ suturing device both ex vivo and in vivo. In the ex-vivo studies, 12-Fr trocar defects were created with a fetoscope in five specimens of human uterine tissue with fetal membranes attached. Specimens were examined for integrity of the anchoring stitch. For the in-vivo studies, trocar defects were created in the two uterine horns of three pregnant ewes, each carrying twins at approximately 79-90 days' gestation. One trocar defect in each ewe was repaired using the suture device, and the other was left unrepaired as a control. The repair sites were examined for membrane-anchoring integrity when the defect was created and at delivery., Results: Fetal membranes were anchored successfully to the uterine myometrium using the suture-delivery device in all five experiments performed ex vivo. The in-vivo experiments also revealed successful membrane anchoring compared with controls, both at the time of device deployment and 1-9 weeks after the procedure., Conclusions: We successfully anchored amniotic membranes to the underlying myometrium using a suturing device, both ex vivo and in vivo. Further studies are needed to evaluate the efficacy of the device and to determine whether it can successfully anchor fetal membranes percutaneously in human patients. © 2024 International Society of Ultrasound in Obstetrics and Gynecology., (© 2024 International Society of Ultrasound in Obstetrics and Gynecology.)

There are several reasons for skin damage, including genetic factors, disorders, acute trauma, hard-to-heal wounds, or surgical interventions. Whatever the cause, wounds have a substantial impact on people who experience them, their caregivers and the healthcare system. Advanced wound care products have been researched and developed, providing an opportunity for faster and more complete healing. Tissue engineering (TE) is a promising strategy that can overcome limitations when choosing a graft for a wound. Amniotic membrane is a highly abundant, readily available, and inexpensive biological tissue that does not raise ethical concerns, with many applications in different fields of TE and regenerative medicine. It has attractive physical characteristics, such as elasticity, rigidity and mechanical strength, among others. The effects can also be potentiated by association with other substances, such as hyaluronic acid and growth factors. This paper describes new perspectives involving the use of amniotic membranes.

Background: Diabetic foot ulcers (DFUs) constitute a complication that occurs in 19% to 34% of patients with diabetes mellitus (DM). The aim of this study is to describe median days to healing, average velocity of wound closure, and percentage of wound surface closed at 3, 6, and 12 weeks through the use of homogenized and lyophilized amniotic membrane (hAMpe) dressings for the treatment of DFUs in ambulatory patients., Methods: An observational, descriptive, longitudinal study was performed. Patients presenting with granulation-based DFU, after proper debridement, were included from August 19, 2021, until July 14, 2023. hAMpe dressings placed every 3 days were used for the treatment of these ulcers., Results: Sixteen patients were included with a mean age of 52.38 (8.07) years. The analyzed lesions were postsurgical ulcers in 15 of the 16 included patients. Median ulcer size was 19.5 cm 2 (6.12-36). The median ABI was 1.10 (1-1.14). The median days to healing was 96 (71-170). The median percentage closure of the wound at 3 weeks was 41% (28.9%-55.3%), at 6 weeks it was 68.2% (48.6%-74.2%), and at 12 weeks it was 100% (81%-100%). The average velocity closure was 1.04% per day (95% CI 0.71%-1.31%). It was higher during the closure of the first 50% of the ulcer, 2.12% per day (95% CI 0.16%-4.09%), and decreased from 50% to 25% of the ulcer size to 0.67% per day (95% CI 0.23%-1.10%) and from 25% to closure to 0.47% per day (95% CI 0.14%-0.80%), P  < .001., Conclusion: These results are difficult to compare to other studies given the higher surface area of the ulcers included in our sample. The development of hAMpe dressings enables patients to apply them without requiring assistance from health care teams and was not associated with any recognized complications., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: Mariano Berra, BPharm, reports non-financial interests from AmniosBMA Tissue Bank (non profit organization), as Technical Director. Alejandro Berra, PhD, reports participation on a data safety monitoring board or advisory board from AmniosBMA Tissue Bank (non profit organization), as R&D Director. Disclosure forms for all authors are available online.

Purpose: Chronic radiation cystitis (CRC) develops after radiation therapy and can present with symptoms like urinary frequency, urgency, pelvic pain, and nocturia. We have previously reported that amniotic bladder therapy (ABT) provides symptomatic improvement in refractory CRC patients for up to 3 months. Herein, we evaluated the durability of ABT up to 6 months., Materials and Methods: CRC patients recalcitrant to previous treatments received ABT comprised of intra-detrusor injections of 100 mg micronized AM diluted in 10 mL 0.9% preservative-free sodium chloride. Clinical evaluation and questionnaires (Interstitial Cystitis Symptom Index (ICSI), Interstitial Cystitis Problem Index (ICPI), Bladder Pain/Interstitial Cystitis Symptom Score (BPIC-SS), Overactive Bladder (OAB) Assessment Tool, SF-12 Health Survey) were repeated at pre-op and 2, 4, 8, 12, 16, 20, 24, and 36 weeks post-injection., Results: Five consecutive patients with a mean age of 64.4 ± 20.1 years with a median CRC duration of 10 years were included and followed for 6 months. After ABT, the lower urinary tract symptoms improved as early as 2 weeks and were maintained up to 20 weeks. BPIC significantly improved from 36.6 ± 1.1 at baseline to 12.6 ± 1.5 at 16 weeks and 13.8 ± 2.9 at 20 weeks. At 24 and 36 weeks, the improvement was maintained in four (80%) of the five patients (BPIC = 13.8 ± 1.0). Uroflow assessment showed voiding volume improved two-fold in four of the five patients at 24 weeks compared to baseline., Conclusion: Our data suggest that a significant number of CRC patients may have durable benefit after ABT. Despite this, some of them can show symptoms rebound at 24 weeks., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objective: The application of canine amniotic membrane (cAM) for corneal reconstruction is widely used in the veterinary field. However, the information on biological properties and alternative forms of cAM for corneal wound healing is limited. This study aimed to investigate the proteomic profiles and corneal wound healing properties of cAM, cAM extract (cAME), and lyophilized cAM extract (cAMX)., Animal Studied: A total number of 14 cAMs were sterilely harvested from healthy full-term puppies and randomly divided into three different forms: cAM (n = 14), cAME (n = 14), and cAMX (n = 14)., Procedures: Each form of cAMs was subjected to proteomic analysis using label-free liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), followed by bioinformatic analysis. The proteins were classified into properties by comparing them with the literature search on human amniotic membrane (hAM) properties and the effect on corneal wound healing when given topically., Results: The analyses identified 8136 proteins in cAM, 8211 proteins in cAME, and 7093 proteins in cAMX. A total number of 100 proteins were matched with proteins in hAM properties and were classified into anti-inflammatory, anti-fibrotic, anti-microbial, anti-angiogenic, promotion of epithelialization, analgesic, and support cell adhesion and growth properties. Furthermore, proteins with corneal wound healing effects were identified in cAME and cAMX., Conclusions: cAM and its extracts contain numerous proteins, including proteins related to corneal wound healing properties. Additionally, cAME and cAMX showed proteins involved in corneal wound healing and their potential benefits for topical use in ophthalmology., (© 2024 American College of Veterinary Ophthalmologists.)

Premature rupture of membranes (PROM) is defined as rupture of fetal membranes before the onset of labor. Prolactin (PRL) is secreted by decidual membranes and accumulated significantly in the amniotic fluid during pregnancy. PRL could ameliorate inflammation and collagen degradation in fetal membranes. However, the role of PRL in amniotic membrane is not well characterized. We isolated human amniotic epithelial stem cells (hAESCs) from human fetal membranes to study the effect of PRL on proliferation, migration, and antioxidative stress. Amniotic pore culture technique (APCT) model was constructed to evaluate the tissue regeneration effect in vitro. The potential targets and pathways of PRL acting in amnion via integrated bioinformatic methods. PRL had a dose-dependent effect on hAESCs in vitro. PRL (500 ng/mL) significantly improved the viability of hAESCs and inhibited cell apoptosis, related to the upregulation of CCN2 expression and downregulation of Bax, Caspase 3, and Caspase 8. PRL accelerated migration process in hAESCs via downregulation of MMP2, MMP3, and MMP9. PRL attenuated the cellular damage and mitochondrial dysfunction induced by hydrogen peroxide in hAESCs. PRL accelerated the healing process in the APCT model significantly. The top 10 specific targets (IGF1R, SIRT1, MAP2K1, CASP8, MAPK14, MCL1, NFKB1, HIF1A, MTOR, and HSP90AA1) and signaling pathways (such as HIF signaling pathway) were selected using an integrated bioinformatics approach. PRL improves the viability and antioxidative stress function of hAESCs and the regeneration of ruptured amniotic membranes in vitro. Thus, PRL has great therapeutic potential for prevention and treatment of ruptured membranes., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com. See the journal About page for additional terms.)

Background: The perfect repair of damaged skin has always been a constant goal for scientists; however, the repair and reconstruction of skin is still a major problem and challenge in injury and burns medicine. Human amniotic membrane (hAM), with its good mechanical properties and anti-inflammatory, antioxidant and antimicrobial benefits, containing growth factors that promote wound healing, has evolved over the last few decades from simple skin sheets to high-tech dressings, such as being made into nanocomposites, hydrogels, powders, and electrostatically spun scaffolds. This paper aims to explore the historical development, applications, trends, and research hotspots of hAM in wound healing., Methods: We examined 2660 publications indexed in the Web of Science Core Collection (WoSCC) from January 1, 1975 to July 12, 2023. Utilizing bibliometric methods, we employed VOSviewer, CiteSpace, and R-bibliometrix to characterize general information, identify development trends, and highlight research hotspots. Subsequently, we identified a collection of high-quality English articles focusing on the roles of human amniotic epithelial stem cells (hAESCs), human amniotic mesenchymal stem cells (hAMSCs), and amniotic membrane (AM) scaffolds in regenerative medicine and tissue engineering., Results: Bibliometric analysis identified Udice-French Research Universities as the most productive affiliation and Tseng S.C.G. as the most prolific author. Keyword analysis, historical direct quotations network, and thematic analysis helped us review the historical and major themes in this field. Our examination included the knowledge structure, global status, trends, and research hotspots regarding the application of hAM in wound healing. Our findings indicate that contemporary research emphasizes the preparation and application of products derived from hAM. Notably, both hAM and the cells isolated from it - hADSCs and hAESCs are prominent and promising areas of research in regenerative medicine and tissue engineering., Conclusion: This research delivers a comprehensive understanding of the knowledge frameworks, global dynamics, emerging patterns, and primary research foci in the realm of hAM applications for wound healing. The field is rapidly evolving, and our findings offer valuable insights for researchers. Future research outcomes are anticipated to be applied in clinical practice, enhancing methods for disease prevention, diagnosis, and treatment., (© 2024 The Author(s). Skin Research and Technology published by John Wiley & Sons Ltd.)

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 activities highlighting the impact of processing method on the versatility of clinical use of amniotic membrane allografts.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., Competing Interests: Declaration of conflicting interestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: All authors were full time employees at MIMEDX Group, Inc. at the time experiments were executed. S.M., M.M., H.B., T.K., and J.H. hold equity in MIMEDX Group, Inc.

Limbal stem cell deficiency (LSCD) is a very challenging situation and difficult to manage. A great works and ideas were conducted over the past 50 years. Numerous surgical techniques were proposed. We are reporting more than a 20-year follow-up of a case of limbal autograft stem cell transplantation due to LSCD secondary to chemical injury. [ABSTRACT FROM AUTHOR]

Surgical repair of severe rotator cuff tear often results in retear due to unaddressed muscle degeneration. The objective of this study was to test the regenerative potential of micronized dehydrated Human Amnion/Chorion Membrane (dHACM), in a clinically relevant delayed reattachment model of rotator cuff repair. Micronized dHACM was injected into rat supraspinatus muscle during tendon re-attachment surgery, three weeks after original tendon injury. One week after material injection, inflammatory and mesenchymal stem cell infiltration into supraspinatus muscles was assessed via flow cytometry. Histological methods were utilized to assess structural and regenerative changes in muscle one and three weeks after material injection. Micronized dHACM injection resulted in increased M1-like macrophages (17.1 × fold change over contralateral controls) and regenerating muscle fibers (4.3% vs 1.7% in saline treated muscles) one week after injection compared to saline treated muscles. Tendon reattachment itself exhibited intrinsic healing in this model, demonstrated by a general return of muscle weight and reduced fibrosis. Our results indicate that injection of micronized dHACM may initiate an inflammatory response in degenerated muscle that promotes early muscle regeneration, and that our animal model may be a suitable platform for studying treatments in muscle at early timepoints, before intrinsic healing occurs. [ABSTRACT FROM AUTHOR]

Objective: This study was designed to fabricate a suitable permanent scaffold for the normal aligned myotube formation and improve the process of myogenic differentiation of selected stem cells. Materials and Methods: In this experimental study, an engineered scaffold composed of decellularized human amniotic membrane (DHAM) and electrospun fibers of poly(ε-caprolactone) (PCL) was fabricated and characterized. PCL nanofibers were superimposed on DHAM (PCL-DHAM) in two different patterns, including randomized fibers (Random) and aligned fibers (Aligned). Adipose derived stem cells (ADSCs) were isolated from adult Wistar rats and cultured on designed scaffold and induced to myotube differentiation. Using an MTT assay, the vitality of cells was determined. Then, myogenic cell differentiation was assessed using scan electron microscopy (SEM), immunofluorescence assay, and reverse transcription-polymerase chain reaction (RT-PCR). Results: The mechanical properties of engineered PCL-DHAM composite improved significantly compared to DHAM as a control. The engineered PCL-DHAM promoted cell growth and high expression of myosin, Mhc2 and myogenin and thus enhanced the myotube formation. Conclusion: These findings revealed that bio-composite membrane prepared from PCL nanofibers and DHAM, may represent a promising biomaterial as a desirable scaffold for applying in the bioengineered muscle repair. [ABSTRACT FROM AUTHOR]

Background: It is now well established that factors (free or in extracellular vesicles) secreted by mesenchymal stromal cells (MSC) are important mediators of MSC regenerative actions. Herein we produced the secretome (conditioned medium, CM) from MSC isolated from the amniotic membrane (hAMSC) and CM from the intact amniotic membrane (hAM, no manipulation or enzymatic digestion) in order to potentially identify an effective, easy and less expensive secretome to produce for potential applications in regenerative medicine. Given that immunomodulation is a key mechanism of action through which hAMSC contributes to tissue regeneration, we used a comprehensive panel of in vitro immunomodulatory tests to compare the CMs. Methods: Amniotic membranes were either cut into fragments or used for hAMSC isolation. CMs from hAMSC at passages 0 and 2 were collected after a standard 5-day culture while CM from hAM was collected after a 2- and 5-day culture. Immunomodulation was assessed in terms of PBMC and T-cell proliferation, T-cell subset polarization, T-regulatory cell induction, cell cytotoxicity and monocyte differentiation toward antigen-presenting cells. Furthermore, we performed a comparison between CM obtained from single donors and pooled CM. We also assessed the impact of lyophilization on the immunomodulatory properties of CM. Results: We demonstrate that CM from hAM has comparable immunomodulatory properties to CM from hAMSC at passages 0 and 2. Furthermore, we demonstrate that pooled CMs have similar effects when compared to CM from single donors used separately. Finally, we demonstrate that lyophilization does not alter the in vitro immunomodulatory properties of CM from hAM and hAMSC. Conclusions: The results presented herein support the possibility to produce secretome from intact hAM and open the prospect to highly improve the scalability of the GMP production process while reducing the costs and time related to the process of cell isolation and expansion. Moreover, the possibility of having a lyophilized secretome that maintains its original properties would allow for a ready-to-use product with easier handling, shipping and storage. The use of a lyophilized product will also facilitate clinicians by permitting customized reconstitution volumes and methods according to the most suitable formula required by the clinical application. [ABSTRACT FROM AUTHOR]

Previously, we reported a collagenase-based, animal product-free protocol for cultivated oral mucosal epithelial cell sheets for transplantation (COMET). Here, we reported the long-term outcomes of first 2 clinical cases. A 27-year-old man suffered from thermal burn, which resulted in symblepharon of lower fornix OD. COMET was performed, and the cornea remained clear with few peripheral NV and no more symblepharon 34 months postoperatively. Another 42-year-old man suffered from severe alkaline burn OD. He underwent COMET, followed by corneal transplantation half a year later. A biopsy taken two years after COMET showed stratified epithelium positive for keratin 4, 13, and 3 in the suprabasal layer. Staining for p63 and p75NTR was both positive in the basal layer. The graft remained clear up to post-OP 4 years. Our study confirmed the long-term survival of the transplanted OMECs, suggesting that collagenase-based spheroidal suspension culture is a promising technique for COMET. Trial registration ClinicalTrials.gov, ClinicalTrials.gov ID: NCT03943797 Registered 9 May 2019-Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT03943797. [ABSTRACT FROM AUTHOR]

Recent reviews suggest that amniotic membrane products may accelerate healing of diabetic foot ulcers. A new dried human amniotic membrane (dHAM) has been used for ocular ulcers but not for diabetic foot ulcers. This was a multi‐centre, prospective, patient and observer blind, randomised controlled pilot trial, to investigate whether 2 weekly addition of the dHAM to standard care versus standard care alone increased the proportion of healed participants' index foot ulcers within 12 weeks. Thirty‐one people (mean age 59.8 years, 81% male, 87% type 2 diabetes) were randomised (15 dHAM, 16 usual care). Within 12 weeks, healing occurred in 4 (27%) ulcers in the dHAM group versus 1 (6.3%) usual care group (P =.1). Percentage wound area reduction was higher in the dHAM versus control group. (P =.0057). There was no difference in AEs between the two groups. Six participants allocated to dHAM correctly identified their treatment group, although 5 in usual care incorrectly thought they were in the intervention arm. This pilot trial result is encouraging showing that this dHAM preparation is safe and promising treatment. These results will be used to design a statistically powered, definitive double blind randomised controlled trial. [ABSTRACT FROM AUTHOR]

Background: The amniotic membrane (AM) is used in ocular surface reconstruction and is effective at promoting epithelialization and preventing corneal perforation in cases of acute microbial keratitis. Here, we report a case of isolated AM infection after AM transplantation for a persistent epithelial defect following tectonic penetrating keratoplasty. Case presentation: A 47-year-old man with poorly controlled type 2 diabetes mellitus presented to the emergency department with a referral for perforated microbial keratitis. After ophthalmic examination, corneal scraping was performed, and corneal gluing was attempted and failed. Hence, the patient underwent tectonic penetrating keratoplasty. After keratoplasty, the patient developed a persistent epithelial defect that required AM transplantation as an overlay. Thirty days post-AM transplant, the patient presented with signs and symptoms resembling granular microbial infection of the cornea. After two days, the granular findings began dislodging from the corneal surface and were sent for culture, sensitivity, and histopathological identification. Histological analysis of the granular material indicated it to be a small piece of AM stroma infiltrated with mixed-type inflammatory cells. Corneal scraping cultures indicated Streptococcus mitis and Streptococcus oralis. Conclusion: The infiltrate was localized to the basement membrane of the AM as, despite the anti-inflammatory effects of AM, it can also act as a barrier against polymorphonuclear leukocyte infiltration from the tear film and microbial invasion into the cornea.

Objective: Inflammation, catabolism, and hypertrophy in chondrocytes play a central role in osteoarthritis (OA). The Wnt/β-catenin and NF-κβ pathways contribute to these degradative processes. This in vitro study evaluates the inhibitory effect of a novel therapeutic, micronized dehydrated human amnion/chorion membrane (μdHACM), as a potential treatment to offset elevated Wnt/β-catenin and NF-κβ signaling. Design: Three-dimensional human articular chondrocyte pellets were stimulated with an inflammatory cocktail to induce a degenerative phenotype. Treatments included varying doses of μdHACM. Protein and gene expression were analyzed using qRT-PCR, immunoblotting, and immunofluorescence to assess changes in the major constituents of Wnt/β-catenin and NF-κβ signaling. Regulation of catabolic activity was evaluated using enzymatic assays that detect MMP-13 and aggrecanase-mediated degradation products in conditioned media. Results: Confirmation of the model was established through the expression of specific markers and extracellular matrix genes, verifying a chondrogenic phenotype was maintained. Inflammatory stimulation elicited a change in the chondrocyte proteome and secretome, elevating Wnt/β-catenin and NF-κβ signaling and downstream expression of inflammatory, proteolytic, and hypertrophic markers, while decreasing primary cartilage matrix components, ACAN and COL2A1. μdHACM reversed these inflammatory-induced changes, suppressing phospho-GSK-3β, β-catenin expression/nuclear localization of the Wnt signaling axis and inhibiting IKKβ, phospho-IκBα, and phospho-p65 in the NF-κβ signaling cascade. Additionally, μdHACM altered expression of direct downstream targets, namely MCP1, MMP3, MMP13, ADAMTS4, ADAMTS5, RUNX2 and COL10A1. Moreover, μdHACM reduced MMP-13 and aggrecanase-mediated substrate degradation. Conclusion: μdHACM ameloriated the effects of inflammatory-induced degeneration in chondrocytes through Wnt/β-catenin and NF-κβ inhibition, subsequently downregulating key inflammatory, hypertrophic and catabolic mediators in vitro.

Objectives: Repairing articular cartilage damage is challenging. Clinically, tissue engineering technology is used to induce stem cell differentiation and proliferation on biological scaffolds to repair defective joints. However, no ideal biological scaffolds have been identified. This study investigated the effects of amniotic membrane/collagen scaffolds on the differentiation of adipose-derived stem cells (ADSCs) and articular cartilage repair.Methods: Adipose tissue of New Zealand rabbits was excised, and ADSCs were isolated and induced for differentiation. An articular cartilage defect model was constructed to identify the effect of amniotic membrane/collagen scaffolds on cartilage repair. Cartilage formation was analyzed by imaging and toluene blue staining. Knee joint recovery in rabbits was examined using hematoxylin and eosin, toluidine, safranine, and immunohistochemistry at 12 weeks post-operation. Gene expression was examined using ELISA, RT-PCR, Western blotting, and immunofluorescence.Results: The adipose tissue was effectively differentiated into ADSCs, which further differentiated into chondrogenic, osteogenic, and lipogenic lineages after 3 weeks’ culture in vitro. Compared with platelet-rich plasmon (PRP) scaffolds, the amniotic membrane scaffolds better promoted the growth and differentiation of ADSCs. Additionally, scaffolds containing the PRP and amniotic membrane efficiently enhanced the osteogenic differentiation of ADSCs. The levels of COL1A1, COL2A1, COL10A1, SOX9, and ACAN in ADSCs + amniotic membrane + PRP group were significantly higher than the other groups both in vitro and in vivo. The Wakitani scores of the ADSC + amniotic membrane + PRP group were lower than that in ADSC + PRP (4.4 ± 0.44**), ADSC + amniotic membrane (2.63 ± 0.38**), and control groups (6.733 ± 0.21) at week 12 post-operation. Osteogenesis in rabbits of the ADSC + amniotic membrane + PRP group was significantly upregulated when compared with other groups. Amniotic membranes significantly promoted the expression of cartilage regeneration-related factors (SOX6, SOX9, RUNX2, NKX3-2, MEF2C, and GATA4). The ADSC + PRP + amniotic membrane group exhibited the highest levels of TGF-β, PDGF, and FGF while exhibiting the lowest level of IL-1β, IL6, and TNF-α in articular cavity.Conclusion: Amniotic membrane/collagen combination-based scaffolds promoted the proliferation and cartilage differentiation of ADSCs, and may provide a new treatment paradigm for patients with cartilage injury.

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.

Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) is a spectrum of rare, severe immunological blistering skin reactions which are triggered by medication intake or infections. The acute phase is characterized by necrolysis of the skin and desquamation of mucosa, primarily oral and ocular, with significant mortality rates. The chronic phase is characterized by multi-organ sequelae with increased rates of morbidity and reduced quality of life for patients who have survived the acute phase. Since the primary goal in the acute phase is saving the life of the patient, ocular involvement is often missed and a significant proportion of patients present to an ophthalmologist with the chronic ocular sequelae. In India, chronic ocular sequelae and low vision are observed in two-thirds of patients who present in the chronic phase of SJS/TEN. In the chronic phase of ocular involvement, there are definite windows of opportunity which if targeted with specific interventions such as scleral lenses and mucous membrane grafts can help reduce the incidence of corneal blindness and improve the quality of life for patients with SJS/TEN. Over the last decade, several studies from India have advanced the understanding of the natural course of ocular involvement in SJS/TEN and the outcomes of timely interventions in the chronic phase of the disease. We present an overview of the epidemiology of ocular complications of SJS/TEN in India, the specific challenges faced in the management of ocular complications in the acute stage and recent advances in management of the chronic ocular complications of the disease.

The application of amniotic membrane (AM) on chronic wounds has proven very effective at resetting wound healing, particularly in re-epithelialization. Historically, several aspects of AM effect on wound healing have been evaluated using cell models. In keratinocytes, the presence of AM induces the activation of mitogen-activated protein (MAP) kinase and c-Jun N-terminal kinase (JNK) pathways, together with the high expression of c-Jun, an important transcription factor for the progression of the re-epithelialization tongue. In general, the levels of transforming growth factor (TGF)-β present in a wound are critical for the process of wound healing; they are elevated during the inflammation phase and remain high in some chronic wounds. Interestingly, the presence of AM, through epidermal growth factor (EGF) signaling, produces a fine-tuning of the TGF-β signaling pathway that re-conducts the stalled process of wound healing. However, the complete suppression of TGF-β signaling has proven negative for the AM stimulation of migration, suggesting that a minimal amount of TGF-β signaling is required for proper wound healing. Regarding migration machinery, AM contributes to the dynamics of focal adhesions, producing a high turnover and thus speeding up remodeling. This is clear because proteins, such as Paxillin, are activated upon treatment with AM. On top of this, AM also produces changes in the expression of Paxillin. Although we have made great progress in understanding the effects of AM on chronic wound healing, a long way is still ahead of us to fully comprehend its effects.

Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) can cause significant damage to the ocular surface and eyelids. The sloughing and inflammation of the ocular mucosal epithelium during the acute phase may lead to scarring sequelae of the eyelids and ocular surface, resulting in pain and vision loss. Amniotic membrane transplantation (AMT) to the eyes and eyelids during the initial 1–2 weeks of the disease can decrease the chronic sequelae. The main development in the ophthalmologic treatment of SJS/TEN in the USA over the last 15 years has been the use of AMT on the ocular surface and eyelids during the acute phase. The evolution of AMT techniques, refinement of the evaluation of the eyes in acute SJS, and the efforts to increase the use of AMT in the USA are discussed.

The preparation of unique wet and dry wound dressing products derived from unprocessed human amniotic membrane (UP-HAM) is described. The UP-HAM was decellularized, and the constituent proteins were cross-linked and stabilized before being trimmed and packed in sterile Nucril-coated laminated aluminium foil pouches with isopropyl alcohol to manufacture processed wet human amniotic membrane (PW-HAM). The dry type of PD-HAM was prepared by decellularizing the membrane, UV irradiating it, lyophilizing/freeze-drying it, sterilizing it, and storing it at room temperature. The UP-HAM consists of a translucent yellowish mass of flexible membranes with an average thickness of 42 μm. PW-HAM wound dressings that had been processed, decellularized, and dehydrated had a thinner average thickness of 30 μm and lacked nuclear-cellular structures. Following successful decellularization, discrete bundle of fibrous components in the stromal spongy layers, microvilli and reticular ridges were still evident on the surface of the processed HAM, possibly representing the location of the cells that had been removed by the decellularization process. Both wet and dry HAM wound dressings are durable, portable, have a shelf life of 3–5 years, and are available all year. A slice of HAM dressing costs 1.0 US$/cm2. Automation and large-scale HAM membrane preparation, as well as storage and transportation of the dressings, can all help to establish advanced technologies, improve the efficiency of membrane production, and reduce costs. Successful treatment of wounds to the cornea of the eye was achieved with the application of the HAM wound dressings. The HAM protein analysis revealed 360 μg proteins per gram of tissue, divided into three main fractions with MWs of 100 kDa, 70 kDa, and 14 kDa, as well as seven minor proteins, with the 14 kDa protein displaying antibacterial properties against human pathogenic bacteria. A wide range of antibacterial activity was observed after treatment with 75 μg/ml zinc oxide nanoparticles derived from human amniotic membrane proteins (HAMP-ZnO NP), including dose-dependent biofilm inhibition and inhibition of Gram-positive (S. aureus, S. mutans, E. faecalis, and L. fusiformis) and Gram-negative bacteria (S. sonnei, P. aeruginosa, P. vulgaris, and C. freundii).

Background and Objectives: Regarding the increasing spread of bacterial resistance, researchers are always interested in finding effective antibiotics of natural origin. The amniotic membrane and blood platelet concentrate are two biological products with an antibacterial effect. The present study aimed to investigate the antibacterial effect of the biological products on broad-spectrum MBL-producing Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. Subjects and Methods The amniotic membrane, blood platelet concentrate, S. aureus, and P. aeruginosa isolates were collected from hospitals in Gorgan City, Iran. The isolates were identified using the biochemical tests. The methicillin-resistant S. aureus and MBL-producing P. aeruginosa strains were collected by the combined disk and iodometric methods. The antibacterial effects of platelet serial dilutions of bacteria were prepared using 0.5 McFarland turbidity standard suspension in tubes. Then, different concentrations of bacteria were mixed with platelet. After four different encounter durations, a sample was obtained and cultured on medium and bacterial growth was examined. The amniotic membrane was assessed by disk diffusion methods. Results The results showed that all isolates of P. aeruginosa and S. aureus were MBL producers. The platelet concentrate showed the antibacterial effect on all S. aureus isolates, whereas it lacked such an effect on P. aeruginosa isolates. It indicates that the amniotic membrane has an antibacterial effect on all S. aureus and P. aeruginosa isolates. Conclusion The amniotic membrane and platelet concentrate showed high antimicrobial potential against multidrug-resistant S. aureus and P. aeruginosa pathogens. Therefore, human-derived natural products can be used as a source for efficient antibiotics. [ABSTRACT FROM AUTHOR]

Background: Paediatric burns are a major health concern, and treatment of choice for less extensive intermediate-depth scald burns is debatable. In a developing country like India where the incidence and complications are high, the need for an easy and cost-effective treatment is highly necessary. The study aimed to retrospectively analyse the need for interventions in paediatric scald burns in a single tertiary referral centre over 4 years, comparing early application amniotic membrane vs wet collagen and paraffin gauze dressings. Methods: All paediatric scald burn patients admitted to the Department of Plastic and Reconstructive Surgery between Jan 2015 and Dec 2018 were identified. Demographics were collected from hospital records and operation theatre logbook. Acute scald burns in patients younger than 15 years were analysed according to dressing applied to find the need for intervention, days of hospitalisation, antibiotics usage and rate of infection. Results: A total of 71 paediatric scald burn patients were analysed between 2015 and 2018. Thirty-six were treated with early application of human amniotic membrane and the remaining 35 were treated with wet collagen and paraffin gauze dressings. Patients treated with human amniotic membrane had lower in-hospital stay (p 0.000) and underwent less frequent dressing change (p 0.046), and none of them needed grafting (p 0.000). Conclusions: Our study results suggest that early application of human amniotic membrane can significantly reduce the need for surgical intervention and should be the preferred method of treatment in less extensive paediatric scald burns. Level of evidence: Level III, therapeutic study. [ABSTRACT FROM AUTHOR]

Background: Heart failure is one of the most common cardiovascular disorders and is considered a chronic, progressive and debilitating disorder. The medical treatment of this disease is accompanied by many problems. Today, stem cells are being used increasingly to reduce the problems of heart failure treatments. Since pro-inflammatory cytokines play an important role in the prognosis and progression of cardiovascular disease, the present study aimed to investigate the effect of intravenous injection of human amniotic membrane mesenchymal stem cells on the levels of interleukins 4 and 12 in the serum of male rats in the heart failure model. Methods: This is an experimental study that was conducted from October 2018 to May 2019 in the Physiology Research Center of Iran University of Medical Sciences. In this study, 28 male wistar rats (180-200 gr) were randomly divided into four groups: control group, heart failure group, heart failure group that received culture medium and heart failure group that received mesenchymal stem cells by intravenous injection. After 30 days, echocardiography was done and then serum levels of interleukin 4 and 12 were measured in these groups by Elisa test . Results: The results of this study showed that intravenous injection of human amniotic membrane mesenchymal stem cells into male rats with heart failure, improved echocardiographic parameters such as ejection fraction (EF) and fractional shortening (FS) in the cell injection group compared to the heart failure group (P<0.05). Also, the levels of inflammatory cytokines IL-4 and IL-12 were significantly reduced in the cell injection group compared to rats with the heart failure group (P<0.05). Conclusion: Due to the improvement of cardiac parameters and the reduction level of inflammatory cytokines in this study, it seems that human amniotic membrane mesenchymal stem cells play an important role in improving heart failure by reducing the level of inflammation. [ABSTRACT FROM AUTHOR]

Introduction: Corneal limbal stem cell deficiency is a multifaceted, sight-threatening disease process that can be challenging to diagnose and treat. Areas covered: This review discusses normal corneal limbal stem cell health, causes and diagnosis of limbal stem cell deficiency, and the medical and surgical management of limbal stem cell deficiency. Expert opinion: Recently established consensus guidelines for the diagnosis and management of limbal stem cell deficiency will help improve the diagnosis, management, and collaborative study design regarding this disease. When medical management fails, a variety of autologous and allogenic limbal stem cell transplant techniques may be tried. However, these treatments potentially have high morbidity and may be short-lived. Newer strategies of ex vivo cell culture and keratoprosthesis implants may help improve visual outcomes in patients faced with bilateral limbal stem cell deficiency. [ABSTRACT FROM AUTHOR]

Gingival Recession is a common, complex, and unpleasant clinical condition. It is more commonly associated with root hypersensitivity, impaired esthetics and root caries. Successful treatment outcome becomes challenging in such cases. Coronally advanced flap for root coverage is most commonly used technique. Combination of biological membranes derived from human body itself along with coronally advanced flap yields better results. This case report describes a case of bilateral Class I gingival recession treated with amniotic membrane on one side and PRF membrane along with Coronally advanced flap on other side. [ABSTRACT FROM AUTHOR]

• The properties of the amniotic membrane (AM) are ideal for its use as a dural substitute. • AM patches can be processed and stored at low costs and used without impacting operative time. • AM create a good separation plane between the dura mater, cerebral cortex, and myocutaneous layer. • AM pre and postoperative outcomes are the same as biological substitutes ones. A dura mater substitute in decompressive craniectomies must protect the brain while providing a dissection plane between the cortex and myocutaneous layer. The human amniotic membrane (AM) has anti-inflammatory, wound healing, and differentiation properties. We tested AM properties as a dural substitute by comparing the outcomes to biological ones. We prospectively collected data on 25 patients who randomly underwent decompressive craniectomy with lyophilized AM patches and 25 in which biological substitutes were utilized between 2015 and 2019. The AM was laid with the epithelial side facing the brain because of the anti-adhesive proprieties, while the chorion facing the myocutaneous flap. We collected data on demographics, neurological status, comorbidities, and surgical outcomes. Additionally, we created a score – dura mimicking score– and reviewed postoperative imaging and pathological specimens. The majority (96%) of AM grafts were integrated into native dura. Thirteen patients scored as excellent and 11 good on our "dura mimicking score", showing tissue integration ability but no cerebral cortex adhesion. The histopathological analysis showed that AM had thick plates of dense fibrous tissue with small reactive vessels, reactive fibroblasts, and lymphocytes infiltrate. The AM group's first outcomes were not different from the biological substitute patients but higher integration rate to the dura and less adhesion to the myocutaneous flap in AM patients. We documented the anti-adhesive, protective, and integrative properties of AM dural substitute patches in patients who underwent decompressive craniectomies, comparing the intraoperative differences and postoperative outcomes to biological dural substitutes. [ABSTRACT FROM AUTHOR]

• A microgel consisting of decellularized amniotic membrane is proposed to treat radiation-induced lung injury. • The microgels that preserve the structural integrity of the contained cargo can be inhaled into lung for topical treatment. • The microgels can switch the microenvironment from pro-inflammation status to pro-healing status. Radiation-induced lung injury (RILI) is one of the major complications in patients exposed to accidental radiation and radiotherapy for thoracic malignancies. However, there is no reliable approach to effectively treat RILI. With favorable biological and physical characteristics, decellularized extracellular matrix (dECM) from amniotic membrane can not only act as a reactive oxygen species scavenger to attenuate the damage of oxidative stress, but also release bio-factors to polarize the macrophages from M1-like phenotype to M2-like phenotype, thus switching the microenvironment from pro-inflammation status to pro-healing status. Meanwhile, as a promising drug delivery system, microgels with a small size and high surface/volume ratio can not only be inhaled into lung tissues with a desired concentration, but also preserve the structural integrity and the functionalities of the contained therapeutics. Therefore, it will be a potent strategy to combine microgels and amniotic dECM for the treatment of RILI, in which the inhaled microgels show a high surface area to rapidly remove reactive oxygen species and release cargos to reduce the inflammation-mediated damage by modulating macrophage phenotypes. We hypothesize that amniotic dECM microgels will be a promising and effective biomaterial for the attenuation of RILI. [ABSTRACT FROM AUTHOR]

• AdipoQ and visfatin are more expressed in the allantoic that in amniotic membranes. • Extra-embryonic annexes AdipoQ and visfatin are higher expressed in layer than broiler. • AdipoQ and visfatin are absent in amniotic fluid at ED7 in layer than broiler. • AdipoQ and visfatin in amniotic fluid are positively correlated with embryo weight. • In ovo injection of visfatin antibodies increases embryo mortality in layer and broiler. Some evidence showed differences between layer and broiler embryo development. We recently showed that two adipokines, adiponectin and visfatin are expressed in the extra embryonic membranes and fluids. However, their role in the embryo development is unknown. Thus, our objectives were 1. to compare the expression of AdipoQ and its receptors AdipoR1 and AdipoR2 and visfatin in extra-embryonic annexes in broiler and layer breeders during the embryo development and 2. to investigate the role of two adipokines in embryo development in both broiler and layer breed after in ovo injection of blocking antibodies against chicken adiponectin or visfatin. We found that adiponectin, AdipoR1, AdipoR2 and visfatin were mainly more expressed in the allantoic that in amniotic membranes. In addition, these expressions increased according the stage of embryo development. We observed a higher expression in layer than in broiler of AdipoQ in allantoic membranes at ED14 and ED18, of AdipoR1 and AdipoR2 in both allantoic and amniotic membranes at ED7 and ED14 and of visfatin only in allantoic membrane from ED7 to ED18. AdipoQ and visfatin were absent in amniotic fluid at ED7 but present at ED14 or ED18 where higher concentrations were detected in layer than in broiler. Interestingly, we showed a strong positive correlation between Adipo and visfatin concentration in amniotic fluid and the body weight of embryo in both breeds. However, after in ovo injection of Adipo antibodies we did not observe any effect on the embryo mortality whereas injection of visfatin antibodies increased in a dose dependent manner the embryo mortality in both breeds. Taken together, Adipo and visfatin are higher expressed in layer than broiler in extra-embryonic membranes and amniotic fluid. Our data suggest also that visfatin could be a main regulator of embryo development. [ABSTRACT FROM AUTHOR]

Purpose: To describe a novel technique of amniotic membrane (AM) patch graft in the management of double chamber treatment after big-bubble deep anterior lamellar keratoplasty (DALK). Case Report: A 35-year-old male patient with advanced keratoconus underwent big-bubble DALK. Manual lamellar dissection was done due to failed big-bubble. First-day postoperative double chamber was detected. Air bubbling and SF6 injection were tried without any success. Double chamber resolved by fixation of AM transplantation patch graft (1 × 1 mm) over the Descemet's membrane perforation with fibrin glue. Conclusion: Amniotic membrane patch graft can be used in the management of double chamber after DALK not responsive to intracameral gas injection.

Nowadays the development of novel biomaterials which would promote tissue lesions regeneration is one of the crucial tasks of regenerative medicine. In the current research we developed a biological active membrane composed of decellularized, lyophilized human amniotic membrane in combination with growth factors of platelet-rich plasma (PRP) and hyaluronic acid. On the basis of histological examination, scanning electron microscopic examination and DNA content analyses it was drawn a conclusion that Sodium Dodecyl Sulfate (SDS) (Sigma) solution and 1% Triton X-100 solution completely remove all existing cells of placenta and chorionic-amniotic membrane without impairment of their structure. After the processes of decellularization and lyophilization the DNA content was less than 2%. The PRP growth factors as the components biological matrix provide faster regeneration. The latter are fixed on the surface of cellular structure of decellularized, lyophilized human amniotic membrane by the reservoir of gelatinous hyaluronic acid

Mario Martelloni,1 Giulia Montagner,2 Diletta Trojan,2 Roberta Abate3 1Private Dental Practice, Roma 00179, Italy; 2Treviso Tissue Bank Foundation Onlus, Treviso, 31100, Italy; 3Private Dental Practice, Roma 00132, ItalyCorrespondence: Diletta TrojanFondazione Banca dei Tessuti di Treviso Onlus, Piazzale Ospedale, 1, Treviso 31100, ItalyTel +39 0422 322951Fax +39 0422 322849Email dtrojan@fbtv-treviso.orgAbstract: Mucogingival surgery has the objective to obtain coverage of the recession, with a favorable long-term prognosis, but also to minimize postoperative problems and surgery-related risks. Human amniotic membrane is increasingly employed for periodontal tissue repair in order to promote epithelialization and to reduce pain and scar tissue formation. A 38-year-old female patient reports dental hypersensitivity resulting from gingival recession affecting tooth 4.1. A mucogingival surgical procedure was proposed and a partial-thickness flap of epithelial-connective tissue was harvested from the palate and grafted on to the receiving site. The amniotic membrane was positioned at the donor site to reduce postoperative morbidity and to encourage rapid palatal healing. One week after the application of HAM (human amniotic membrane), the wound was healed and 1 month after the donor site was completely re-epithelialized. The present case report suggests that in the surgical treatment of gingival recession with palatal epithelial-connective tissue graft, HAM promotes rapid epithelialization of the palatal donor site wound with a reduction in morbidity.Keywords: gingival recession, amniotic membrane, palatal epithelial-connective tissue, human tissue

Investigation of the superiority of the usage of propolis extract, amniotic membrane transplantation, corticosteroid and antibiotic both separately and in different combinations was aimed. A total of 40 (20 male, 20 female) New Zealand breed rabbits weighing 2.5-3.0 kg were used in the study. The rabbits were divided into 5 groups on day 7 of alkali burn injury. Amniotic membrane transplantation was applied to 3 groups and each one of these 3 groups had different local medications as 1% water-based propolis extract (AMN+PRP), dexamethasone and tobramycin (AMN+AC) and only tobramycin (AMN+A). The other two groups, which were not performed amniotic membran transplantation, were either treated with propolis (PRP) or left as a control. The defect area results obtained from AMN+PRP group were lower than AMN+A group, and both AMN+PRP and control group at the day 9 and day 14, respectively (P≤0.05). In the measurements made after the second week, AMN+PRP group showed significantly better results in all weeks in terms of defect area (P≤0.05). Corneal thickness was found to be lower in all groups treated with amniotic membrane than in the other groups (P

Limbal epithelial stem cells are localized in the limb, which determine the regeneration of the corneal epithelium, both physiological and reparative. The aim of the study is to characterize the culture of cells isolated by different methods.Materials and methods. Limbal stem cells were isolated from the limbus region of enucleated human eyes. Cell isolation was produced by the enzymatic and explantation methods. Cell cultivation was performed on the amniotic membrane and without it, in DMEM/F12 growth media (Dullbecco’s Modified Eagle Medium/Nutrient F12 Ham) and CECBM (Corneal Epithelial Cell Basal Medium). The morphological assessment and immunophenotyping of cell cultures were performed.Results and its discussion. The obtained culture was distinguished by cell heterogeneity at the enzymatic method of isolating cells from the limbus. The presence of fibroblast-like cells was revealed in the suspension of non-adherent cells, which resembled stromal cells in appearance and growth pattern. Immunophenotyping revealed that most cells carry markers of multipotent mesenchymal stromal cells (CD90+/CD73+/CD105+/CD34–/CD45–) and a small number – markers of epithelial cells (p63α+, ABCG2 +, CK19+). Explant culture was more uniform. The cells had a rounded, epithelial-like shape, adhered well to the surface of the amniotic membrane and grew on it. The results of phenotyping indicated that cells are represented mainly by epithelial cells (p63α+, ABCG2 +, CK19+).Conclusion. The culture is represented mainly by MMSC at the enzymatic method of isolating cells from the limbal graft and cultivating them without feeder in the growth medium DMEM/F12. Isolation of limb cells by the explantation method and their cultivation in the selective medium CECBM allow us to obtain mainly the culture of epithelial stem cells.

Abstract Background Skin burn wound is a notable medical burden worldwide. Rapid and effective treatment of burnt skin is vital to fasten wound closure and healing properly. Amniotic graft and Aloe vera are widely used as wound managing biomaterials. Sophisticated processing, high cost, availability, and the requirement of medics for transplantation limit the application of amnion grafts. We aim to prepare a novel gel from amnion combined with the Aloe vera extract for burn wound healing which overcome the limitations of graft. Methods Two percent human amniotic membrane (AM), Aloe vera (AV) and AM+AV gels were prepared. In vitro cytotoxicity, biocompatibility, cell attachment, proliferation, wound healing scratch assays were performed in presence of the distinct gels. After skin irritation study, second-degree burns were induced on dorsal region of Wistar rats; and gels were applied to observe the healing potential in vivo. Besides, macroscopical measurement of wound contraction and re-epithelialization; gel treated skin was histologically investigated by Hematoxylin and eosin (H&E) staining. Finally, quantitative assessment of angiogenesis, inflammation, and epithelialization was done. Results The gels were tested to be non-cytotoxic to nauplii and compatible with human blood and skin cells. Media containing 500 μg/mL AM+AV gel were observed to promote HaCaT and HFF1 cells attachment and proliferation. In vitro scratch assay demonstrated that AM+AV significantly accelerated wound closure through migration of HaCaT cells. No erythema and edema were observed in skin irritation experiments confirming the applicability of the gels. AV and AM+AV groups showed significantly accelerated wound closure through re-epithelialization and wound contraction with P

Elizabeth Yeu,1 Damien F Goldberg,2 Francis S Mah,3 Kenneth A Beckman,4,5 Jodi I Luchs,6,7 Jonathan D Solomon,8 Darrell E White,9 Preeya K Gupta101Eastern Virginia Medical School, Virginia Eye Consultants, Norfolk, VA, USA; 2Wolstan & Goldberg Eye Associates, Torrance, CA, USA; 3Scripps Clinic Torrey Pines, La Jolla, CA, USA; 4Ohio State University, Columbus, OH, USA; 5Comprehensive Eye Care of Central Ohio, Westerville, OH, USA; 6Hofstra Northwell School of Medicine, Hempstead, NY, USA; 7South Shore Eye Care, Wantagh, NY, USA; 8Bowie Vision Institute, Bowie, MD, USA; 9Skyvision Centers, Westlake, OH, USA; 10Duke University Eye Center, Durham, NC, USAPurpose: Evaluate the safety and efficacy of cryopreserved amniotic cytokine extract (ACE) in the treatment of subjects with dry eye disease (DED).Patients and methods: This was a retrospective, multicenter, chart review of adult patients with DED that instilled cryopreserved ACE drops twice-daily for 4 or 12 weeks. Patients had corneal fluorescein staining (0–20 range) and/or a lissamine green conjunctival staining score (0–18 range) of ≥3 and ≤9 in at least 1 eye and a score ≥40 (0–100 range) of eye dryness/irritation on a visual analog scale (VAS). Following completion of a treatment course, medical records were reviewed from the initiation of therapy (baseline), and at post-treatment visits (4 weeks, 8 weeks, and 12 weeks). Patient records for visual acuity, adverse events, corneal fluorescein staining, conjunctival lissamine green staining, and symptom scores of ocular dryness/irritation were reviewed for each visit, as available. Safety and tolerability were assessed through the evaluation of patient-reported adverse events recorded in the medical records.Results: A total of 54 eligible patients were identified at 7 clinical sites; 16 patients administered ACE drops for 4-weeks, and 38 patients instilled ACE drops for 12 weeks. Significant improvements in the mean changes from baseline were observed for corneal fluorescein staining, lissamine green staining, visual acuity (LogMar) and VAS ocular symptom scores at the 4-week post-treatment visit (p