Application Of Human Amniotic Membrane Research Articles

Cuidados de las zonas donantes de injertos de piel de espesor parcial. Umbrella review

ObjectivesTo summarize the current evidence and develop recommendations on the most effective management of partial-thickness graft donor sites in adults to improve re-epithelialization, pain, and infection. MethodologyUmbrella review. A systematic search of Pubmed, CUIDEN, Cochrane Library, CINHAL Plus, SCOPUS, and LILACS databases was conducted between October and December 2021, including systematic reviews published in the last 10 years that inquired about the effectiveness of any partial-thickness graft donor site healing strategy. Reviews with very low critical appraisal according to AMSTAR 2 were excluded. Included reviews were scored using the SIGN scale to determine their level of evidence and grade of recommendation. ResultsFive systematic reviews with meta-analysis were included. A statistically significant improvement in re-epithelialization and pain was observed in platelet-rich plasma and human amniotic membrane in relation to the control group. In addition, wound infection also decreased in platelet-rich plasma. A decrease in re-epithelialization time was also observed in recombinant human growth hormone. ConclusionsThe use of platelet-rich plasma is recommended to improve re-epithelialization, pain, and infection in partial-thickness graft donor sites in adults. The application of human amniotic membrane is recommended to accelerate re-epithelialization and reduce pain, and recombinant human growth hormone is recommended to accelerate the time to completely heal these wounds.

Healing techniques for split-thickness skin grafts donor sites. Umbrella review

ObjectivesTo summarize the existing evidence and provide recommendations for the most effective management of partial-thickness graft donor sites in adults, with the goals of enhancing re-epithelialization, reducing pain, and preventing infection. MethodologyUmbrella review. A systematic search was conducted encompassing databases such as Pubmed, CUIDEN, Cochrane Library, CINHAL Plus, SCOPUS, and LILACS. The search targeted systematic reviews published since 2011 that focused on examining the effectiveness of different approaches for the treatment of partial-thickness graft donor sites. Reviews with a low critical appraisal score according to AMSTAR 2 were excluded. The included reviews were evaluated using the SIGN scale to assess the level of evidence and grade the recommendations. ResultsFive systematic reviews with meta-analysis were incorporated in the analysis. Platelet-rich plasma and human amniotic membrane demonstrated statistically significant improvements in re-epithelialization and pain reduction when compared to the control group. Moreover, platelet-rich plasma also exhibited a decrease in wound infection rates. Recombinant human growth hormone was found to expedite the re-epithelialization process. ConclusionsBased on the findings, the use of platelet-rich plasma is recommended to enhance re-epithelialization, alleviate pain, and reduce infection in partial-thickness graft donor sites among adults. Application of human amniotic membrane is recommended to accelerate re-epithelialization and alleviate pain, while recombinant human growth hormone is suggested to expedite the overall healing time of these wounds.

The Use of Human Amniotic Membrane (hAM) as a Treatment Strategy of Medication-Related Osteonecrosis of the Jaw (MRONJ): A Systematic Review and Meta-Analysis of the Literature.

Background and objectives: Although it is very uncommon, medication-induced osteonecrosis of the jaw (also known as MRONJ) can have serious consequences. Traditionally, this adverse event has been recognised in patients who were treated with bisphosphonate (BP) drugs. Nevertheless, in recent years, it has been established that individuals having treatment with various types of medications, such as a receptor activator of nuclear factor kappa-Β ligand inhibitor (denosumab) and antiangiogenic agents, have had the same issue. The purpose of this research is to determine if the application of human amniotic membrane (hAM) may be used as a therapy for MRONJ. Material and Methods: A multi-source database (MEDLINE, EMBASE, AMED, and CENTRAL) systematic search was performed. The major objective of this study is to obtain an understanding of the efficacy of hAM when it is employed as a treatment modality for MRONJ. The protocol of this review was registered in the INPLASY register under the number NPLASY202330010. Results: The authors were able to include a total of five studies for the quality analysis, whereas for the quantity evaluation, only four studies were eligible. A total of 91 patients were considered for the investigation. After treatment with human amniotic membrane (hAM), a recurrence of osteonecrosis was observed in n = 6 cases (8.8%). The combined efficacy of surgical therapy and the use of hAM resulted in an overall success rate of 91.2%. Intraoperative complications were only documented in one article, and they were mostly caused by the positioning of the hAM, which led to wound breakdown at the surgical site. Conclusions: Based on the small amount of data and low-quality research included in this study, using human amniotic membranes to treat MRONJ might represent a feasible option. Nevertheless, further studies with a wider patient population are required to understand the long-term impacts.

The Use of Amniotic membrane in Carpal Tunnel Syndrom, a Cadaver Study

Introduction: Carpal tunnel syndrome (CTS) is the most common peripheral nerve entrapment syndrome worldwide. Even after surgery in 2-25% of patients symptoms persist. Most commonly the carpal tunnel was not completely released. In the other cases perineural scarring is causing recurrent compression of the nerve. The literature states different treatment options including “nerve wraps,” made out of autologous veins or synthetic materials, to protect the nerve from scarring. The human amniotic membrane (HAM) has been proven to express anti-inflammatory and anti-fibrotic properties, without causing a immune reaction. In a rat model it has been shown that nerve gliding could be improved by the application of HAM after introducing a fibrotic scar. The application of HAM in CTS and it´s possible influence on nerve gliding has been examined. Materials and Methods: Fourteen (7 female, 7 male) fresh body donors were dissected. The carpal tunnel was released and a nerve wrap out of HAM was sutured around the median nerve with continuous stitches (7-0). The position of the HAM was measured and the skin was closed (3-0) Next the arm was moved according to a protocol, creating a largest possible movement of the nerve. The position was again evaluated. This was done after 25, 50, 100, 150, 200, 250 movement cycles. Results: The HAM on the median nerve did not change its position significantly between the timepoints. From the beginning to the end of the movements it contracted and reduced its size to about 92% of its starting length. The evaluation of the gliding properties is still in progress. Conclusion: The HAM showed to stay in place and only minor contract on both ends in the carpal tunnel. The application of HAM in the carpal tunnel worked well without any complications and can be seen as feasible treatment method for recurrent carpal tunnel syndrome.

Human amniotic membrane application in oral surgery—An ex vivo pilot study

Objectives: The purpose of this pilot porcine study was to explore and illustrate the surgical application of human amniotic membrane (hAM) in an ex vivo model of medication-related osteonecrosis of the jaw (MRONJ). Material and methods: Five oral and maxillofacial surgeons participated to this study. MRONJ was simulated on porcine mandible specimens. hAM was applied using four different techniques: implantation with complete coverage, implantation with partial coverage, apposition and covering graft material. At the same time, the surgeons evaluated how well the hAM handled and its physical properties during the surgery. Results: Surgeons found that hAM had suitable mechanical properties, as it was easy to detach from the support, handle, bind to the defect and bury. hAM was also found to be strong and stable. The "implantation with complete coverage" and "implantation with partial coverage" techniques were the preferred choices for the MRONJ indication. Conclusion: This study shows that hAM is a graft material with suitable properties for oral surgery. It is preferable to use it buried under the gingiva with sutures above it, which increases its stability. This technical note aims to educate surgeons and provide them with details about the handling of hAM in oral surgery. Clinical relevance: Two surgical techniques for hAM application in MRONJ were identified and illustrated. hAM handling and physical properties during surgery were reported.

Sıçanlarda Mastektomi ve Aksiller Diseksiyon Sonrası Seroma Gelişimi Üzerine Kriyoprezerve İnsan Amniyotik Membran ve Trombositten Zengin Plazmanın Etkileri

Aim: Seroma is one of the most common complications after mastectomy and axillary dissection. It occurs as a result of prolongation of the exudative-inflammatory phase of wound healing. The aim of this study is to evaluate the effects of human amniotic membrane (HAM) and platelet-rich plasma (PRP) on seroma formation. Material and Methods: A total of 24 rats were grouped as control, PRP, and HAM groups. All rats underwent radical mastectomy and axillary lymph node dissection. Saline in the control group, PRP in the second group, and HAM in the third group were applied to the dissection area. The groups were compared in terms of the condition of the surgical field, seroma volume, and histopathological changes. Results: Seroma volume decreased in the PRP group, but not in the HAM group. Lymphocyte, eosinophil, histiocyte, and fibroblast levels were significantly lower both in the PRP and HAM groups compared to the control group. E-cadherin and TGF-β1 immunoreactivities of PRP and HAM groups were higher than in the control group. Conclusion: In this study, it was observed that PRP and cryopreserved HAM increased tissue healing and decreased the inflammatory process. However, although local PRP application significantly reduced seroma formation, it was determined that HAM application did not reduce seroma formation. It was thought that this might be due to the fact that the smooth surface of HAM mechanically prevents the adhesion of the tissues. More beneficial results will be obtained with the modification to be made in the preparation and application of HAM.

Surgical Application of Human Amniotic Membrane and Amnion-Chorion Membrane in the Oral Cavity and Efficacy Evaluation: Corollary With Ophthalmological and Wound Healing Experiences.

Due to its intrinsic properties, there has been growing interest in human amniotic membrane (hAM) in recent years particularly for the treatment of ocular surface disorders and for wound healing. Herein, we investigate the potential use of hAM and amnion-chorion membrane (ACM) in oral surgery. Based on our analysis of the literature, it appears that their applications are very poorly defined. There are two options: implantation or use as a cover material graft. The oral cavity is submitted to various mechanical and biological stimulations that impair membrane stability and maintenance. Thus, some devices have been combined with the graft to secure its positioning and protect it in this location. This current opinion paper addresses in detail suitable procedures for hAM and ACM utilization in soft and hard tissue reconstruction in the oral cavity. We address their implantation and/or use as a covering, storage format, application side, size and number, multilayer use or folding, suture or use of additional protective covers, re-application and resorption/fate. We gathered evidence on pre- and post-surgical care and evaluation tools. Finally, we integrated ophthalmological and wound healing practices into the collected information. This review aims to help practitioners and researchers better understand the application of hAM and ACM in the oral cavity, a place less easily accessible than ocular or cutaneous surfaces. Additionally, it could be a useful reference in the generation of new ideas for the development of innovative protective covering, suturing or handling devices in this specific indication. Finally, this overview could be considered as a position paper to guide investigators to fulfill all the identified criteria in the future.

Early application of human amniotic membrane in paediatric scald burns decreases the need for surgical intervention

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. 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. 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). 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.

Use of Human Amniotic Membrane to Achieve Permissive Hypovolaemia in Burn Patients

In thermal burn patients, epidermis and dermis are damaged and the barrier function of the skin is lost. This leads to fluid, nutrients and heat loss along with the increased risk of microbial contamination. Both inadequate resuscitation and over resuscitation are deleterious to health. The increasing trend of overhydrating patients is well reported, and various guidelines to prevent this are in practice. Given its easy availability and cost-effectiveness, the human amniotic membrane can be used as a temporary barrier dressing. The aims of this study are (1) to evaluate the efficacy of amniotic membrane in decreasing the amount of replacement fluid in burn patients and (2) to measure the difference of fluid requirement by application of amniotic membrane compared with Parkland formula. Patients admitted to our institution between December 2016 and January 2018, who underwent early burn blister debridement with human amniotic membrane application, were included in this study. In this case-control study of thermal burn patients (including both adults and children), comparisons were made between historical patients from hospital record who also had equivalent burns and were managed by conventional prevailing dressing (cotton gauze dressings) against prospectively collected patients in whom entire burn surface was completely covered with human amniotic membrane dressing. The intravenous requirement in the historical group was generally based on Parkland formula calculation and strict urinary output maintenance. In the prospective group, the desired urinary output was maintained, and accordingly intravenous fluid was titrated to the necessary minimum. The fluids administered in the first 24 and 48 h were compared against the fluid requirement as per the Parkland formula. In our study, we found a mean reduction of 25.52% requirement of resuscitation fluid in the first 24 h after application of the human amniotic membrane. Our study provides evidence that the application of human amniotic membrane allows us to successfully resuscitate burn victims with lesser intravenous fluid than prescribed by Parkland formula. Human amniotic membrane dressing can be a good addition to colloid infusion in preventing fluid creep.

<p>Amniotic Membrane In Palatal Epithelial-Connective Tissue Reconstruction</p>

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.

Benefits of cryopreserved human amniotic membranes in association with conventional treatments in the management of full-thickness burns.

The use of split-thickness skin autografts (STSA) with dermal substitutes is the gold standard treatment for third-degree burn patients. In this article, we tested whether cryopreserved amniotic membranes could be beneficial to the current treatments for full-thickness burns. Swines were subjected to standardised full-thickness burn injuries, and then were randomly assigned to treatments: (a) STSA alone; (b) STSA associated with the dermal substitute, Matriderm; (c) STSA plus human amniotic membrane (HAM); and (d) STSA associated with Matriderm plus HAM. Clinical and histological assessments were performed over time. We also reported the clinical use of HAM in one patient. The addition of HAM to classic treatments reduced scar contraction. In the presence of HAM, skin wound healing displayed high elasticity and histological examination showed a dense network of long elastic fibres. The presence of HAM increased dermal neovascularization, but no effect was observed on the recruitment of inflammatory cells to the wound. Moreover, the use of HAM with classical treatments in one human patient revealed a clear benefit in terms of elasticity. These results give initial evidence to consider the clinical application of HAM to avoid post-burn contractures and therefore facilitate functional recovery after deep burn injury.

Development of a visible light, cross-linked GelMA hydrogel containing decellularized human amniotic particles as a soft tissue replacement for oral mucosa repair.

Early effective treatment of oral mucosal defects is the key to ensuring defect healing and functional recovery. The application of human amniotic membrane (HAM) in promoting wound healing has been shown to be safe and effective. However, amniotic membrane is thin, easy to tear and difficult to handle. Combined with the natural forces at play in the oral cavity, this has restricted the clinical applications of HAM for healing of mucosal defects. Methacrylated gelatin (GelMA) has good mechanical strength and adhesion, and can be used as a bionic repair film to attach to the damaged surface of oral mucosa, but GelMA lacks bioactive substances and cannot promote the rapid repair of oral mucosal defects. The aim of this study was to design a type of composite GelMA hydrogel mixed with decellularized human amniotic particles (dHAP) as an oral mucosa substitute, to promote regeneration of defective mucosa by stimulating rapid angiogenesis. The composite substitute GelMA–dHAP was easy to synthesize and store, and easy to operate for repair of oral mucosal defects. We show the angiogenic potential of GelMA–dHAP on chick chorioallontoic membrane and the curative effect of GelMA–dHAP as a treatment in the rabbit oral mucosa defect model. In conclusion, this study confirms the effectiveness of GelMA–dHAP as an ideal soft tissue substitute for the repair of oral mucosal defects, overcoming the shortcomings of using HAM or GelMA alone.

Application of amniotic membrane in reconstructive urology; the promising biomaterial worth further investigation

ABSTRACTIntroduction: In reconstructive urology, autologous tissues such as intestinal segments, skin, and oral mucosa are used. Due to their limitations, reconstructive urologists are waiting for a novel material, which would be suitable for urinary tract wall replacement. Human amniotic membrane (AM) is a naturally derived biomaterial with a capacity to support reepithelization and inhibit scar formation. AM has a potential to become a considerable asset for reconstructive urology, i.e., reconstruction of ureters, urinary bladder, and urethrae.Areas covered: This review aims to discuss the potential application of human AM in reconstructive urology. The environment for urinary tract healing is particularly unfavorable due to the presence of urine. Due to its fetal origin, the bioactivity of AM is orientated to induce intrinsic regeneration mechanisms and inhibit scarring. This review introduces the concept of applying human AM in reconstructive urology procedures to improve their outcomes and future tissue engineering based strategies.Expert opinion: Many fields of medicine that have accomplished translational research have proven the usefulness of AM in clinical practice. There is an urgent need for studies to be conducted on large animal models that might convincingly demonstrate the underestimated potential of AM to urologists around the world.

Human Amniotic Membrane Improves Healing in a Chronic, Massive Rotator Cuff Repair Model

Objectives:Rotator cuff tendon heals by fibrovascular scar that is weaker than native tissue leading to repairs that are prone to failure. Our objective was to investigate the ability of an amniotic membrane-derived human allograft to improve rotator cuff tendon-bone healing and skeletal muscle architecture in a chronic massive rotator cuff injury and repair model in rats. We hypothesized that application of human amniotic membrane to the tendon-bone interface at the time of repair will result in increased attachment strength and rotator cuff muscle fiber force secondary to improved bone formation and nthesis histomorphometry, less plasminogen activator inhibitor-1 (PAI-1) expression, and induced muscle repair gene expression.Methods:Eighty-five male Sprague-Dawley rats were allocated to one of four groups: (1) uninjured (U), (2) injury only (IO), (3) chronic injury and repair (CR), or (4) chronic injury and augmented repair (ER). The IO, CR, and ER groups underwent unilateral detachment of the supraspinatus and infraspinatus tendons with injection of the corresponding muscle with 3 u/kg Onabotulinum toxin A. Four weeks following injury, a surgical repair was performed in the CR and ER groups using transosseous suture fixation with human amniotic membrane applied to the tendon-bone repair site in the ER group. Animals were euthanized at four weeks following the repair. Biomechanical testing of the supraspinatus and infraspinatus tendon-bone complex and single fiber contractility testing of the supraspinatus muscle were performed. Microcomputed tomography was utilized to quantitate bone microstructure at the repair site. The healing tendon-bone interface was evaluated with histomorphometric quantification of fibrocartilage formation and collagen organization, and immunostaining with PAI-1. The rotator cuff muscle was evaluated with immunohistochemical localization for lipid deposition and total cross-sectional area. Real-time polymerase chain reaction evaluated relative expression of genes relevant to muscle repair. Statistical analysis was performed using a nonparametric Kruskal-Wallis test with significance set at p<0.05.Results:Augmentation with amniotic membrane did not lead to increased load-to-failure or stiffness compared to CR. The cross-sectional area, maximum isometric force, and specific force of individual supraspinatus muscle fibers were significantly greater with ER compared to CR (Figure 1). Microcomputed tomography revealed a larger volume of newly formed bone present at the bony footprint with ER compared to CR. Histomorphometric analysis demonstrated significantly greater fibrocartilage area and collagen organization at the healing tendon-bone insertion site with ER compared to CR at 4 week. Qualitatively, there was less PAI-1 immunostaining noted at the tendon-bone interface in the ER group. There was less lipid deposition and greater cross-sectional area of the rotator cuff muscle fibers in the ER group. Rotator cuff muscle gene expression with ER demonstrated a downregulation of apoptosis regulatory genes BCL2 and Caspase-3, adipogenesis regulatory genes FITM1 and FITM2, TNF-alpha and TGF-beta 3 compared to CR.Conclusion:Augmentation with human amniotic membrane in a chronic, massive rotator cuff injury and repair model led to greater bone formation, more organized tendon enthesis, induced muscle repair gene expression, and greater rotator cuff muscle fiber force secondary to less PAI-1 expression.

In Vitro Evaluation of the Drug Reservoir Function of Human Amniotic Membrane Using Moxifloxacin as a Model Drug.

To evaluate the in vitro, extended drug reservoir function of human amniotic membrane (HAM) of different thicknesses impregnated with moxifloxacin. HAM buttons (12 mm) were soaked with freshly prepared 0.5% wt/vol topical moxifloxacin at different soaking time intervals: 3 hours (group I), 6 hours (group II), 12 hours (group III), 24 hours (group IV), and 48 hours (group V). They were then transferred into 1 mL of fresh simulated tear fluid (pH-7.4) and incubated at 37°C. The release kinetics of moxifloxacin was studied by analyzing the amount of drug in simulated tear fluid collected at different time intervals from each pretreated HAM for 3 weeks. In another experiment, thin and thick HAMs were selected based on weight and soaked with moxifloxacin for 24 hours, and the release kinetics was studied for 7 weeks. All samples were stored at -80°C until analysis by high-performance liquid chromatography. No significant difference was observed between different soaking times and the release of moxifloxacin. The cumulative amount of moxifloxacin released from thick HAM was found to be statistically significant compared with thin HAM (P < 0.05). Our in vitro data showed that the sustained release of moxifloxacin from HAM was achieved up to 7 weeks. The entrapment efficiency of moxifloxacin was significantly higher in thicker HAM than in thin HAM. Moxifloxacin-impregnated HAM application can be considered in bacterial keratitis to provide sustained drug delivery through a biological bandage system for up to a period of 7 weeks.

Simultaneous application of human amniotic membrane and Tachosil® in the repair of recto-vaginal fistula in an animal model

Patients with recto-vaginal fistula (RVF) suffer from distressing symptoms like the passage of stool and gas through the vagina. Although surgeons employed various techniques to address this disorder, however, current techniques are invasive and their success rate is variable. Considering the notable features of human amniotic membrane (HAM) as a bio-prosthesis and Tachosil® as a type of biologic mesh, the present study was designed to assess the effect of HAM in the presence of Tachosil® on the repair of RVF in a rabbit model. Twelve New Zealand rabbits weighing 2–2.5 kg with the age of 6–8 months were selected, and RVF was created surgically in all of them. They were randomly divided into three groups for repair with HAM, repair with HAM in the presence of Tachosil®, and standard repair, respectively. Histopathological features were evaluated using modified scoring system. Statistical analysis was performed by SPSS software version 18 (SPSS Inc., Chicago, IL, USA) using the Kruskal-Wallis test. Gross examination and microscopic evaluation were performed after 4 weeks. Fistula healing occurred in all rabbits and no abscess or infection was detected. Microscopic fistula healing was scored based on epithelialization, collagenization, inflammation, neovascularization, necrosis, and granulation tissue. Differences among these three groups were not statistically significant (p > 0.05). Our findings revealed that the major positive effect on RVF repair was related to HAM and healing was not improved in the presence of Tachosil®. Further studies with larger sample size would be helpful for more detailed assessment.

Effect of human amniotic membrane on anastomosis leakage in dog model

One of the major concerns in rectal cancer surgery and complicated anastomosis is anastomosis breakdown and leakage. For decades, radiation (as neoadjuvant or adjuv ant therapy) was known as a risk factor for anastomosis leakage. Diverting loop ileostomy has been usually employed to divert the fecal flow and protect complicated anastomosis. However, there is a high rate of complications related to diverting loop ileostomy and its closure. This experimental study was carried out to investigate the effect of human amniotic membrane on colonic anastomosis in dogs that underwent preoperative radiation therapy. In this prospective controlled animal trial study, a total of ten male cross-breed dogs aged 6–8 months and weighing 10–15 kg were randomly assigned in to two groups of five dogs. Group 1 with five dogs received 10 Gy of external radiation preoperatively, while the second didn’t receive radiation. After anesthesia and exploration, about 8 cm above the dentate line, the bowel was cut and end-to-end anastomosis was done in two layers hand sewn technique. HAM measuring 2.5 cm × 2.5 cm was wrapped around the anastomosis and sutured with vicryl 3-0. In each group, one of the dogs died. The non-parametric Mann—Whitney test was used to compare the two groups. Two-tailed P value 0.05, i.e., healing in both irradiated and non-irradiated groups was the same). Unfavorable effects of radiotherapy and loop ileostomy may be resolved with application of human amniotic membrane, which influences anastomosis healing and eliminates the need for ileostomy insertion.

Liver diseases: what is known so far about the therapy with human amniotic membrane?

Liver, the largest intern organ of the human body, is responsible for several vital tasks such as digestive and excretory functions, as well as fornutrients storage and metabolic functions, synthesis of new molecules and purification of toxic chemicals. Cirrhosis, fibrosis and hepatocellular carcinoma are the most prevalent liver diseases. Despite all the studies performed so far, treatment options for these diseases are very limited. For this reason, it is urgent to find effective therapies for these pathologies. Several studies have been performed during the last decade about the possible application of human amniotic membrane in hepatic diseases therapy. Promising results about human amniotic membrane or its derived cells, in vitro and in vivo, applications in fibrosis, cirrhosis and hepatocellular carcinoma were already published. Since it is an attractive study area, it is becoming a dynamic scientific subject. However, the action mechanisms of human amniotic membrane and its derived cells in hepatic diseases therapy must be precisely known in order that this promising therapy could be clinically used.

The amniotic membrane from the human placenta contains different subregions with different morpho-functional features

Human Amniotic Membrane (AM) has been attributed anti-inflammatory, anti-angiogenic, anti-fibrotic and anti-microbial effects (1-2). A number of clinical studies have used AM for applications in regenerative medicine, but it has yet to be elucidated whether the different areas of the AM have the same plasticity and differentiation potential. Thus, the aim of our study was to map the AM from normal human placenta. AMs were obtained from 24 healthy women undergoing caesarean section. Four areas were considered with respect to the umbilical cord: the first area, closer to the umbilical cord, was named the central area; the second, in the middle, was the intermediate area; the third was the peripheral area (all the three areas were part of the chorion frondosum) and the fourth, the reflected area, corresponded to the chorion laeve. Our results demonstrate the presence of a multi-layered epithelium in different areas of AM, except for the intermediate area, with a number of budding or detaching cells as well as of apoptotic cells, especially in the central area. By means of immune-histochemistry and quantitative analysis with MetaMorph analysis software, we evaluated the in situ expression of different proteins observing that the peripheral area has the highest OCT-4, c-KIT and SOX-2 expression, well known indicators of pluripotency, and the highest levels of CREB and p-CREB, well known for its key role in proliferation, differentiation and apoptosis, whereas mainly the central area expresses high levels of α-fetoprotein, suggesting that it could be more prone to hepatic differentiation. Interestingly, amniotic epithelial cells (AECs) from the peripheral area display the highest quantity of granules in the cytoplasm, as shown in electron microscopy preparations. In addition, in vitro cultured AECs derived from the reflected area demonstrate the highest clonogenic capacity, whereas those from the intermediate area show the highest osteogenic potential, when induced to osteogenic differentiation. Our study is the first to propose the morpho-functional mapping of the AM as a useful tool to identify areas with different stemness properties and plasticity. This could increase the efficiency of human AM application within a therapeutic context.This work was supported by funds granted from MIUR to Prof. Roberta Di Pietro and Lucia Centurione and by Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy to Prof. Ornella Parolini.

Biotechnological application of human amniotic membrane in skin reconstruction

The human amniotic membrane (HAM) has a long history of use in surgical diagnostic. Therefore, the discoveries of cell populations from HAM, which are capable to differentiate into different skin types have been stimulating further research in characterizing this cells and their biotechnological application. Since the discovery of HAM-derived cell characteristics, the interest of big pharmaceutical industry has become more visible in application to skin grafts. In this context, we provide an overview of the recent progress and future perspectives of HAM-derived cells in skin tissue reconstruction and further biotechnological application.