Use Of Human Amniotic Membrane Research Articles

Experimental study on the effects of human amniotic membrane in combination with menstrual blood-derived stem cells on wound healing in a diabetic rat model

One of the serious challenges in diabetic patients is the occurrence of complications caused by the disease. One of the most important side effects is wounding in limbs. Due to the multifactorial nature of these wounds, treatments require a multifaceted approach. Therefore, the aim of the present study was whether the human amniotic membrane (HAM) in combination with menstrual blood-derived stem cells (MenSCs) could promote wound healing in diabetic rats. Thirty days after induction of diabetes, the animals were randomly allocated into four equal groups (n=15): the control group, HAM group, MenSC group, and HAM+MenSC group. Sampling was done on days 7, 14, and 21 for histological, molecular, and tensiometrical evaluations. The results showed that the wound healing rate, collagen deposition, volumes of new epidermis and dermis, as well as tensiometrical characteristics were significantly increased in the treatment groups compared to the control group, and these changes were more obvious in the HAM+MenSC ones (P<0.05). Moreover, the expression levels of TGF-β, bFGF, and VEGF genes were considerably increased in treatment groups compared to the control group and were greater in the HAM+MenSC group (P<0.05). This is while expression levels of TNF-α and IL-1β decreased more significantly in the HAM+MenSC group than the other groups (P<0.05). We concluded that the combined use of HAM and MenSCs has a more significant effect on diabetic wound healing.

A pilot study of the use of human amniotic membrane as subcutaneous implants in a mouse model: a potential for temporary substitutes in two-stage breast reconstructions

IntroductionBreast reconstruction by prosthesis is frequently performed in breast cancer treatments, and a temporary substitute is used in the first step of two-stage operations.AimDue to the advantageous biological features of the human amniotic membrane, we aimed to evaluate its use for temporary implants.MethodWe prepared small spherical implants from human amniotic membranes and inserted them into BALB/c mice’s subcutaneous flanks. Then, we compared the bulging they produced, the durability, and the host reaction with implants made from the chorionic membrane, folded membrane patches, and sterile plastic beads.ResultsAll amionitic cases were healthy throughout the study and only mild inflammation occurred in them. Furthermore, the bulging of the implants was acceptable and faded gradually. However, moderate inflammation was observed in chorionic implant mice, and the bulging disappeared very soon. Finally, the control group had severe inflammation and the beads implant was rejected.ConclusionOur study showed that the human amniotic membrane could represent a safe and valid tool for breast reconstruction, however, further studies on larger animals and more implants are suggested.

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.

Comparison of freeze dried human amniotic membrane and its combination with platelet rich fibrin for the epithelial acceleration in an acute wound excision using Rattus Novergicus study model

Link of Video Abstract: https://youtu.be/FvU45HL3HyE Introduction: Human amniotic membrane and platelet rich fibrin (PRF) are biological materials that can accelerate wound healing through their content of growth factors and cytokines. This study aimed to determine the effectiveness of the combination of these two biological agents for wound healing. Methods: This study used an experimental method with a randomized post-test only control group design, to compare the use of human amniotic membrane alone with a combination of human amniotic membrane and PRF in acute full thickness wounds of the skin of Wistar rats. The research subjects were 32 rats which were divided into four groups. Groups 1 and 2 used human amniotic membrane, while groups 3 and 4 used a combination of human amniotic membrane and PRF, then the epithelialization process was observed on the 3rd and 6th day. Result: There was no statistically significant difference in the speed of epithelialization of excision wounds on the backs of rats on the third day (Significance 0.498) and Sixth (Significance 0.9) between the Amnion group and the combination of amnion and PRF because it was greater than α = 0.05. Conclusion: The use of a combination of freeze dried human amniotic membrane and platelet rich fibrin was not significantly more effective than freeze dried human amniotic membrane in accelerating the epithelialization of acute full thickness excision wounds in rats (Rattus novergicus).

Human Amniotic Membrane Usage from Bench to Bedside in Congenital Defects and Wounds in Paediatric Surgery: A Systematic Review

Background: The restricted donor area in paediatric patients demands the use of Human Amniotic membrane (hAM) in the management of difficult-to-manage wounds. It can be used directly over wounds or used to grow stem cells by different culture methods. The hAM can be used “fresh” i.e., +4O glycerol preserved or “cryopreserved”. Methods: In this literature review, we searched ‘PubMed’, ’Web of Science’ and ’SCOPUS’ for experimental models, RCT, Observational studies, case series, and case reports involving the usage of hAM in the treatment of neonatal and paediatric (​&lt;14 yrs.) patients, published in from 1993 to April 2022. The search included the keywords, “amnion”, “biomaterials”, “biological dressing”, “clinical study”, “congenital defects”, “human amniotic membrane”, “paediatric wound”, “neonatal wounds”, and “regenerative medicine”. The Search was extended by snowballing the reference list of all included studies. Results: The final analysis included one experimental RCT, three review articles, and twelve case reports. The experimental studies were in rat, pup, and porcine models. The paediatric second-and third-degree thermal burn followed by paediatric ocular diseases viz corneal epithelial ulcers, conjunctiva reconstruction following Steven Johnson Syndrome, and scarring after surgery of strabismus were the most common indications for the usage of AM. Five cases of meningomyelocele repair (intradural &amp; extradural placement of AM) and 2 cases of gastroschisis repair (as an antiadhesive layer) were reported. Freeze-dried hAM is most frequently used in clinical practice. Autologous hAM was used in antenatally detected birth defects. In the adults, the fresh hAM was found equally effective as freeze-dried AM, but with the risk of transmission of contagious diseases. The literature on Fresh amnion is deficient in paediatric patients. Conclusion: hAM as a skin substitute in paediatric wounds/defects has shown an enhanced rate of healing. However, further studies, regarding the utility of hAM in the management of paediatric wounds, congenital anatomical defects, and diseases along with analysis of outcome and economic constraints in developing countries are needed.

Human amniotic membrane as a dural substitute in neurosurgery: A systematic review

Several studies have highlighted the use of human amniotic membrane (HAM) in neurosurgical procedures as an effective dural substitute. HAM has inherent antifibrotic and anti-inflammatory properties and exhibits immunomodulatory effect that makes it an ideal dural substitute. Other advantages including easy availability, low cost of procurement, and storage also render it a promising dural substitute especially in low- and middle-income countries. A systematic literature search was performed using PubMed, Scopus, and Google Scholar databases, using the search terms "human amniotic membrane," "dural repair," and "neurosurgery." To be eligible for inclusion in our review, papers had to report primary data, be published in English language and report dural repair on humans with human amniotic membrane. Eligibility assessment was conducted by two independent reviewers with qualitative analysis on the basis of surgical utility, postoperative complications, and histological analysis. Eight articles met the predefined inclusion criteria, including three randomized control trials and five cohort studies. We evaluated the use of HAM grafts in dural repair for elective cranial surgery (four studies), trauma surgery (three studies), and elective spine surgery (one study). Cases with postoperative cerebrospinal fluid (CSF) leak were reported by two studies. Other postoperative complications including meningitis, hydrocephalus, pseudomeningocele, CSF collection in subdural space, and subacute subdural hematoma were reported by one study each. Postsurgical histological analysis was reported by three studies highlighting the antiadhesive and integrative properties of HAM. The current review of evidence suggests that in terms of postsurgical outcomes, HAM is comparable with commercially available dural substitutes.

Human Amniotic Membrane Plug for Chronic or Persistent Macular Holes

To report on the use of human amniotic membrane for macular holes at 2 Canadian tertiary care centers. Retrospective cohort study. Twenty-two patients with persistent or chronic macular holes. Macular hole surgery was performed by 3 vitreoretinal surgeons. Macular hole closure with complete plugging by human amniotic membrane on spectral-domain OCT. The closure rate was 91% (20 of 22; median follow-up, 7 months), with no statistically significant visual acuity change overall. Complications included subretinal silicone oil (5%), choroidal neovascularization (5%), atrophy (5%), and cystoid macular edema (9%). Anatomical success with limited visual recovery was observed.

The clinical effectiveness of cryopreserved human amniotic membrane in diabetic foot syndrome

Diabetic foot syndrome with long-term unhealed wounds is the most common complication and cause of limb amputation in diabetes. The search for effective therapeutic agents and their inclusion in treatment protocols is a priority due to the increase in the number of cases of this socially significant disease and disability among the working population every year. The aim of the study is to evaluate the effectiveness of cryopreserved human amniotic membrane in the treatment of long-term non-healing wounds of the lower extremities in diabetic foot syndrome. Materials and methods. The pilot clinical study described 4 clinical cases of treatment of patients with diabetes mellitus type I and II (1 woman and 3 men aged 52 to 68 years) with long-term unhealed wounds of the limbs under standard therapy. After previous wound sanation the applications of the cryopreserved human amniotic membrane were performed. Once a week after the application, the dynamics of wound healing was assessed. Blood glucose levels were determined before amniotic membrane treatment and two hours after the procedure. Results. As a results of weekly applications of human amniotic membrane there was a gradual decrease in the area of the wound from the original size and increase the rate of healing. Thus, at the time of the second visit (after 7 days) the reduction in the area of the ulcer from the initial size in patient 1 was 33 %, patient 2 – 25 %, patient 3 – 33 % on the sole and patient 4 – 3 %, and the healing rate – 4.7 %, 3.6 %, 4.7 % and 0.43 % per day, respectively. The use of human amniotic membrane did not affect blood glucose levels when comparing values before application and two hours after the procedure. Regular follow-up visits of patients 3, 6, 9 and 12 months after the start of the study showed no recurrence of ulcers. Conclusions. It has been shown that the use of cryopreserved human amniotic membrane in patients with diabetes mellitus and diabetic foot syndrome with long-term unhealed wounds results in complete healing of ulcers with stable remission during the year of observation.

Human Amniotic Membrane Plug for Macular Holes Coexisting with Rhegmatogenous Retinal Detachment.

PurposeTo evaluate the efficacy of human amniotic membrane in promoting closure of macular holes coexisting with rhegmatogenous retinal detachment.MethodsThis is a retrospective case series of 14 eyes (14 patients) with macula off retinal detachment. These patients had a macular hole coexistent with peripheral retinal breaks. A human amniotic membrane plug was used to close the macular hole during vitrectomy without ILM peeling.ResultsThe mean preoperative BCVA (logMAR value) was 1.87 ± 0.31. At the 6-month follow-up visit, the mean LogMAR best-corrected visual acuity was 0.67 ± 0.17. At the 6-month follow-up, all patients showed complete retinal reattachment with macular hole closure.ConclusionThe use of human amniotic membrane is a valid option in surgery for macular holes coexisting with rhegmatogenous retinal detachment.

Human amniotic membrane: A biological cost effective dressing in chronic wounds.

Objectives: The use of human amniotic membrane is essential new concept in wound healing which functions as a biodegradable scaffold on wound surface, as it is a rich hub of stem cells which play an important role in wound healing. Study Design: Randomized Control Trial. Setting: Department of Surgery THQ Hospital Gojra. Period: 1st January 2019 to 30 September 2019. Material &amp; Methods: Experimental study using clinical trial. A case series of 50 patient cases were picked from surgical OPD. Who fall in criteria of chronic non-healing wound with at least three months duration comprising of diabetic, venous ulcers and traumatic non healing wound and neuropathic ulcers. All located on lower limbs. Results: All 50 patient were treated with standard protocol by applying freshly prepared amniotic membrane out of which 4 chronic wounds more than 4 year duration were not healed and 2 cases escaped from the study. HAM dressing was changed after every 7 days and its effect were studied by seeing measuring the reduction in wound size and improvement in pain, swelling and mental stress. Success rate was found about 90% with complete healing. Conclusion: There is a dire need in developing countries to promote the use of HAM, in chronic non healing wounds which is a biological membrane, readily available (free if fresh) with simple sterilization techniques, easy storage and easy application with ultimate goal in achieving speedy cost effective wound healing.

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.

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.

Effect of the Mode of Application of Cryopreserved Human Amniotic Membrane on Adhesion Formation after Abdomino-Pelvic Surgery in a Mouse Model.

Adhesions after abdomino-pelvic surgery are a cause of morbidity and reoperations. The use of human amniotic membrane (HAM) for adhesion prevention has given controversial results. The mode of administration of the amniotic membrane has not been well studied. This study assessed the efficacy of two modes of application of cryopreserved HAM, patch or fragmented in Lactated Ringer (LR) solution, for the prevention of pelvic adhesion formation postabdomino-pelvic surgery in a mice model. After a midline laparotomy incision, a small cautery lesion was done on each side of the abdominal wall peritoneum in mice. In Group A (control; n = 42), the abdomen was closed directly, Group B (n = 42) received 2.5 ml of LR prior to closure. In Groups C (n = 42) and D (n = 42), a 2 cm × 2 cm patch of HAM and another one fragmented and dispersed in 2.5 ml of LR were applied prior to closure, respectively. Two weeks later, a laparotomy was performed, and gross and pathological evaluation of adhesions, fibrosis, angiogenesis, and inflammation were conducted. Group D exhibited a significantly lower rate of gross adhesion formation. Fibrosis was significantly lowest in Group C as compared to the control. Group B had the lowest vascular formation in the adhesions. The use of HAM fragmented in LR solution is associated with a significantly lower incidence of postoperative adhesions in mice when compared to LR alone, HAM patch, or control. The mechanism of action of this reduction needs to be elucidated by future studies.

Human Amniotic Membrane Transplantation - Different Modalities of its Use in Ophthalmology

The amniotic membrane (AM) is the inner layer of the fetal membranes and consist of 3 different layers: the epithelium, basement membrane and stroma which further consists of three contiguous but distinct layers: the inner compact layer, middle fibroblast layer and the outermost spongy layer. The AM has been shown to have anti-inflammatory, anti-fibrotic, anti-angiogenic as well as anti-microbial properties. Also because of its transparent structure, lack of immunogenicity and the ability to provide an excellent substrate for growth, migration and adhesion of epithelial corneal and conjunctival cells, it is being used increasingly for ocular surface reconstruction in a variety of ocular pathologies including corneal disorders associated with limbal stem cell deficiency, surgeries for conjunctival reconstruction, as a carrier for ex vivo expansion of limbal epithelial cells, glaucoma surgeries and sceral melts and perforations. However indiscriminate use of human AM needs to be discouraged as complications though infrequent can occur. These include risk of transmission of bacterial, viral or fungal infections to the recipient if the donors are not adequately screened for communicable diseases, if the membrane is not processed under sterile conditions or if storage is improper. Optimal outcomes can be achieved only with meticulous case selection. This review explores the ever expanding ophthalmological indications for the use of human AM.

The effect of human amniotic membrane as a covering layer on propylene mesh in decrease of adhesion after laparotomy in the rabbit

Each year, approximately 4 million abdominal surgeries are performed, and it is estimated that 10–20 % of these patients develop abdominal hernias at the incision point. Currently, the standard method for repair of such hernias is the use of polypropylene mesh, which has complications such as adhesions. Therefore, it is crucial and logical to search for a type of material which can reduce such complications. This study was conducted in order to determine surgical and histological outcomes of polypropylene mesh coated with human amniotic membrane (HAM) in comparison with polypropylene mesh on its own. Ten rabbits were divided into two equal groups, group I and II. A 5-cm incision was made along the abdomen of each rabbit. In group I, a 2 × 3 cm layer of polypropylene mesh was placed under the wall of the abdomen adjacent to the viscera. In group II, the procedure was identical except that the mesh was covered by a 4 × 6 cm layer of HAM too. After 3 weeks, all animals were sacrificed and tissue samples were provided from the abdominal wall and adjacent viscera to evaluate any adhesion and histological changes in each group. Microscopic evaluation was performed for all samples, and no significant statistical difference was found between groups. It seems that the use of HAM as a coating for polypropylene mesh could have a positive effect in reducing adhesions in the abdominal viscera.

Repair of oral mucosal defects with cryopreserved human amniotic membrane grafts: prospective clinical study

The aim of this study was to evaluate the clinical outcome of the surgical repair of oral mucosal defects using cryopreserved human amniotic membrane (HAM) as a graft material. Thirty-four patients with precancerous lesions such as leukoplakia, erythroplakia, and verrucous hyperplasia were included. Fresh amniotic membrane was obtained from women undergoing elective caesarean section; the membrane was cleaned, prepared in antibiotic solutions, and preserved at −80°C. Results suggested that HAM promotes healing and epithelialization without specific complications. Thus we conclude that the use of HAM gives promising results in the repair of post-surgical oral mucosal defects.

Human amniotic membrane transplantation: Different modalities of its use in ophthalmology.

The amniotic membrane (AM) is the inner layer of the fetal membranes and consist of 3 different layers: the epithelium, basement membrane and stroma which further consists of three contiguous but distinct layers: the inner compact layer, middle fibroblast layer and the outermost spongy layer. The AM has been shown to have anti-inflammatory, anti-fibrotic, anti-angiogenic as well as anti-microbial properties. Also because of its transparent structure, lack of immunogenicity and the ability to provide an excellent substrate for growth, migration and adhesion of epithelial corneal and conjunctival cells, it is being used increasingly for ocular surface reconstruction in a variety of ocular pathologies including corneal disorders associated with limbal stem cell deficiency, surgeries for conjunctival reconstruction, as a carrier for ex vivo expansion of limbal epithelial cells, glaucoma surgeries and sceral melts and perforations. However indiscriminate use of human AM needs to be discouraged as complications though infrequent can occur. These include risk of transmission of bacterial, viral or fungal infections to the recipient if the donors are not adequately screened for communicable diseases, if the membrane is not processed under sterile conditions or if storage is improper. Optimal outcomes can be achieved only with meticulous case selection. This review explores the ever expanding ophthalmological indications for the use of human AM.

A new method to repair recto-vaginal fistula: Use of human amniotic membrane in an animal model.

Background:Recto-vaginal fistula is primarily one of the co-morbidities of vaginal delivery. These patients suffer from persistent malodor vaginal discharge. Various surgical techniques have been employed by surgeons in the course of time. This is the first trial of applying Human Amniotic Membrane (HAM) as a bio-prosthesis in repairing recto-vaginal fistula.Materials and Methods:In a prospective animal study, 8 mixed-breed female dogs weighing 23-27 kg with the age of 12-18 months were selected. They were randomly divided into two groups for standard recto-vaginal fistula repair and fistula repair with human amniotic membrane. The Kruskal-Wallis and Mann Whitney tests were performed to indicate statistical differences.Results:After 6 weeks, fistulas were evaluated both grossly and microscopically. In gross examination, there were no difference between the two groups and healing of fistula seemed to have been occurred in all dogs expect for one which had a persistent patent fistulous tract. Microscopic healing was scored according to epithelialization, collagenization inflammation, ulcer and necrosis of samples. Healing score was significantly higher in the HAM group than the standard group (P = 0.029).Conclusion:Our findings revealed that using HAM as a bio-prosthesis to repair recto-vaginal fistula would result in better surgical and histological outcomes comparing to simple repair.

Amniotic membrane in oral and maxillofacial surgery

Following its renaissance in ophthalmology during the 1990s, preserved human amniotic membrane (HAM) has become an attractive biomaterial for all surgical disciplines. This article reviews the current and potential use of HAM in oral and maxillofacial surgery, its postulated properties and common preservation techniques. Literature was identified by an electronic search of PubMed in July 2012; this was supplemented from the reference lists of the consulted papers. HAM has been used in the field of oral and maxillofacial surgery from 1969 onwards because of its immunological preference and its pain-reducing, antimicrobial, mechanical and side-dependent adhesive or anti-adhesive properties. The effects of HAM on dermal and mucosal re-epithelialisation have been highlighted. Typically, HAM is applied after being banked in a glycerol-preserved, DMSO-preserved or freeze-dried and irradiated state. Whereas the use of HAM in flap surgery and in intra-oral and extra-oral lining is reported frequently, novel HAM applications in post-traumatic orbital surgery and temporomandibular joint surgery have been added since 2010. Tissue engineering with HAM is a fast-expanding field with a high variety of future options. Preserved HAM is considered to be a safe and sufficient biomaterial in all fields of oral and maxillofacial wound healing. Recently published novel indications for HAM application lack a high level of evidence and need to be studied further.

Amniotic membrane as part of a skin substitute for full‐thickness wounds: An experimental evaluation in a porcine model

We evaluated the use of human amniotic membrane (HAM) as a graft material for the treatment of iatrogenic full-thickness (FT) skin wounds in a porcine model with a view to reducing donor site morbidity in free flap transfer. Forty experimental FT-wounds were covered with an autologous split-thickness skin graft (STSG) alone or in combination with a mono- or multilayer HAM or Integra(®). Untreated wounds served as controls. Clinical evaluation and biopsy-sampling for histological and immunohistochemical staining with von-Willebrand-factor (vWF) antibody, laminin antibody, Ki-67 antibody, and smooth muscle actin (αSMA) antibody were performed on days 5, 7, 10, 20, 40, and 60 after surgical intervention. Considerable disparities in the estimated criteria were observed between the various treatment groups of the FT-wounds. The use of HAM was found to have an accelerating impact on re-epithelialization. The multilayered amnion membrane showed better results than the Integra(®) and monolayer technique in terms of contraction rate, inflammation, and scarring and seemed useful as a dermal substitute in FT-wounds giving comparable results to STSG coverage alone. This study demonstrates the successful application of HAM as part of a skin substitute in FT-wounds in minipigs. The results offer promise as a simple and effective technique for the application of multilayer HAM in iatrogenic human skin defects and the acceleration of wound healing.