Human Amniotic Membrane Research Articles

Bilateral Crosslinking with Glutaraldehyde and 1-Ethyl-3-(3-Dimethylaminopropyl) Carbodiimide: An Optimization Strategy for the Application of Decellularized Human Amniotic Membrane in Tissue Engineering

Introduction. The decellularized human amniotic membrane (dHAM) emerges as a viable 3D scaffold for organ repair and replacement using a tissue engineering strategy. Glutaraldehyde (GTA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) can increase the biomechanical properties of dHAM. However, the crosslinking process is associated with biochemical changes and residual toxic materials, dampening the biocompatibility of the dHAM. From a histologic point of view, each side of the amniotic membrane is biologically different. While the dHAM basement membrane side is rich in growth factors, the stromal side of the dHAM contains more connective tissue matrix (e.g., collagen fibers) which supports its biomechanical properties. Biocompatibility and biomechanical properties are two important challenges in the field of materials science. In this study, for the first time, the stromal and basement membrane side are cross-linked with GTA and EDC, respectively, to optimize the biocompatibility of the treated dHAM while sparing the GTA-mediated biomechanical improvements. Methods. Crosslinking was carried out on dHAM in three groups: EDC, GTA and bilateral treatment with EDC&GTA. Mechanical resistance, degradability, and crosslinking measurements were performed on treated dHAM. The viability of mesenchymal stem cells (MSCs) on the scaffolds was evaluated by the MTT assay. The expression levels of surface markers and images of the MSCs were thoroughly studied. Results. The results obtained showed that bilateral treatment of dHAM with EDC and GTA increased mechanical resistance. Similarly, the evaluation of surface markers revealed that bilaterally treated dHAM sustains the stemness and viability of MSCs at a level equal to that achieved with EDC alone. The SEM images indicated that the MSCs maintained adhesion on EDC&GTA-cross-linked dHAM. Conclusion. The current study explores a pioneering treatment of dHAM, a material long recognized for its regenerative properties, in a novel context. This research delves into the utilization of dHAM cross-linked with EDC&GTA, demonstrating its optimized efficacy in tissue engineering. The enhanced crosslinking technique significantly alters the membrane’s properties, amplifying its durability and therapeutic potential. In this novel bilateral treatment strategy (EDC and GTA), improving mechanical properties by GTA on the stromal surface and maintaining the biocompatibility of EDC on the side of the basement membrane of dHAM had been attained together. By investigating the handling and impact of this cross-linked membrane, this study unveils a new approach in leveraging a well-known material through an innovative process, revolutionizing its application in wound care.

Introduction to Amniotic Membranes in Maxillofacial Surgery-A Scoping Review.

Background: Amniotic membrane (AM) holds significant promise in various medical fields due to its unique properties and minimal ethical concerns. This study aims to explore the diverse applications of the human amniotic membrane (HAM) in maxillofacial surgery. Methodology: A comprehensive search was conducted on databases, namely Google Scholar, PubMed, and Scopus, from January 1985 to March 2024. Articles in English, Polish, and Spanish were included, focusing on keywords related to amniotic membrane and oral surgery. Results: Various preservation methods for HAM were identified, namely fresh, decellularized, cryopreserved, lyophilized, and air-dried formats. Clinical studies demonstrated the efficacy of HAM in repairing oral mucosal defects, vestibuloplasty, oronasal fistula closure, cleft palate treatment, bone defect repair, and medication-related osteonecrosis of the jaw (MRONJ). Surgeon evaluations highlighted the ease of handling but noted challenges in suturing and stability during application. Conclusions: Amniotic membranes offer a versatile and effective option in maxillofacial surgery, promoting wound healing, reducing inflammation, and providing a scaffold for tissue regeneration. Further research, including randomized trials and comparative studies, is warranted to validate the efficacy and optimize the utilization of HAM in clinical practice.

Histopathological and statistical study of corneal repair in rabbits. A new way to understand corneal recovery.

The study aims to evaluate corneal healing post amniotic membrane transplantation in controlled corneal defects, justifying its application in routine ophthalmology practice. The objective is to establish a reliable method for assessing the repair process. In three groups of six adult New Zealand rabbits, keratectomy and a monolayer transplant of dehydrated human amniotic membrane (AM) were conducted in the left eye (OS) with the right eye (OD) serving as the control eye. Clinical signs were assessed, and both eyes were enucleated at 1, 2, and 4 weeks for optical coherence tomography (OCT) measurements and histological analysis, collecting data from different epithelium, stroma, and limbus regions. This study was conducted using a formula that combines histologic data categorizing their presence and/or type as beneficial for corneal repair. No statistically significant differences were found between the experimental and control eyes regarding all clinical signs and OCT measurements. However, a linear model using histopathological results showed a period-implant mode interaction with statistical significance (p=0.010). The use of the single-layer amniotic membrane resulted in improved corneal recovery with the stromal side showing better performance in the first week and the epithelial side proving to be more effective than the stromal side in the long term. For the first time, a statistical formula employing histopathological data is introduced to determine corneal recovery, potentially offering a more accurate and reliable method compared with the observation of clinical signs and corneal measurements with OCT.

Mesenchymal stem cells derived-exosomes enhanced amniotic membrane extract promotes corneal keratocyte proliferation.

Amniotic membrane extract (AME) and Wharton's jelly mesenchymal stem cells derived-exosomes (WJ-MSC-Exos) are promising therapeutic solutions explored for their potential in tissue engineering and regenerative medicine, particularly in skin and corneal wound healing applications. AME is an extract form of human amniotic membrane and known to contain a plethora of cytokines and growth factors, making it a highly attractive option for topical applications. Similarly, WJ-MSC-Exos have garnered significant interest for their wound healing properties. Although WJ-MSC-Exos and AME have been used separately for wound healing research, their combined synergistic effects have not been studied extensively. In this study, we evaluated the effects of both AME and WJ-MSC-Exos, individually and together, on the proliferation of corneal keratocytes as well as their ability to promote in vitro cell migration, wound healing, and their impact on cellular morphology. Our findings indicated that the presence of both exosomes (3 × 105 Exo/mL) and AME (50 μg/mL) synergistically enhance the proliferation of corneal keratocytes. Combined use of these solutions (3 × 105 Exo/mL + 50 μg/mL) increased cell proliferation compared to only 50 μg/mL AME treatment on day 3 (**** p < 0.0001). This mixture treatment (3 × 105 Exo/mL + 50 μg/mL) increased wound closure rate compared to isolated WJ-MSC-Exo treatment (3 × 105 Exo/mL) (*p < 0.05). Overall, corneal keratocytes treated with AME and WJ-MSC-Exo (3 × 105 Exo/mL + 50 μg/mL) mixture resulted in enhanced proliferation and wound healing tendency. Utilization of combined use of AME and WJ-MSC-Exo can pave the way for a promising foundation for corneal repair research.

Decellularized human amniotic membrane scaffolds: influence on the biological behavior of dental pulp stem cells

ObjectiveThe objective of this study was to assess the characterization of human acellular amniotic membrane (HAAM) using various decellularization methods and their impact on the proliferation and differentiation of human dental pulp stem cells (DPSCs). The goal was to identify scaffold materials that are better suited for pulp regeneration.MethodsSix different decellularization methods were used to generate the amniotic membranes. The characteristics of these scaffolds were examined through hematoxylin and eosin (H&E) staining, scanning electron microscopy (SEM), and immunohistofluorescence staining (IHF). The DPSCs were isolated, cultured, and their capacity for multidirectional differentiation was verified. The third generation (P3) DPSCs, were then combined with HAAM to form the decellularized amniotic scaffold-dental pulp stem cell complex (HAAM-DPSCs complex). Subsequently, the osteogenic capacity of the HAAM-DPSCs complex was evaluated using CCK8 assay, live-dead cell staining, alizarin red and alkaline phosphatase staining, and real-time quantitative PCR (RT-PCR).ResultsOut of the assessed decellularization methods, the freeze-thaw + DNase method and the use of ionic detergent (CHAPS) showed minimal changes in structure after decellularization, making it the most effective method. The HAAM-DPSCs complexes produced using this method demonstrated enhanced biological properties, as indicated by CCK8, alizarin red, alkaline phosphatase staining, and RT-PCR.ConclusionThe HAAM prepared using the freeze-thaw + DNase method and CHAPS methods exhibited improved surface characteristics and significantly enhanced the proliferation and differentiation capacity of DPSCs when applied to them. The findings, therefore demonstrate the capacity for enhanced pulp regeneration therapy.

Acellular human amniotic membrane: A safe and stable matrix for corneal stromal regeneration

Amniotic membrane (AM) transplantation is used as a safe and effective method to treat a variety of ocular complications. On the ocular surface, this membrane degrades within a few days. This study investigated the safety and long-term stability of human amniotic membrane (HAM) grafts in the corneal stroma. Three rabbits with healthy eyes underwent unilateral stromal pocketing with implantation of decellularized HAM (dHAM). Examinations of graft bio-integration were performed by corneal imaging using slit lamp and optical coherence tomography (OCT) at 1 week, 3 months, and 9 months. The thickness of the thickest part of the amniotic membrane was 187 μm at the first follow-up, 184 μm at 3 months, and 166 μm at 9 months. Histological evaluation was performed after the last follow-up. The results showed no clinical complications, such as opacity, neovascularization, infection, or rejection. The thickness of the grafts remained stable over time. Histopathological analyses showed the grafts were integrated into the corneal stroma without any inflammation. This study indicated that HAM was a naturally safe, stable, and compatible matrix in the corneal stroma and was considered a promising candidate for corneal stromal regeneration.

Combined Amniotic Membrane and Self-Powered Electrical Stimulator Bioelectronic Dress Promotes Wound Healing.

Human amniotic membranes (hAMs) are widely used as wound management biomaterials, especially as grafts for corneal reconstruction due to the structure of the extracellular matrix and excellent biological properties. However, their fragile nature and rapid degradation rate hinder widespread clinical use. In this work, we engineered a novel self-powered electronic dress (E-dress), combining the beneficial properties of an amniotic membrane and a flexible electrical electrode to enhance wound healing. The E-dress displayed a sustained discharge capacity, leading to increased epidermal growth factor (EGF) release from amniotic mesenchymal interstitial stem cells. Live/dead staining, CCK-8, and scratch-wound-closure assays were performed in vitro. Compared with amniotic membrane treatment alone, the E-dress promoted cell proliferation and migration of mouse fibroblast cells and lower cytotoxicity. In a mouse full-skin defect model, the E-dress achieved significantly accelerated wound closure. Histological analysis revealed that E-dress treatment promoted epithelialization and neovascularization in mouse skin. The E-dress exhibited a desirable flexibility that aligned with tissue organization and displayed maximum bioactivity within a short period to overcome rapid degradation, implying great potential for clinical applications.

Assessing the Application and Effectiveness of Human Amniotic Membrane in the Management of Venous and Diabetic Ulcers: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

This study aimed to assess the efficacy of human amniotic membranes (HAM) in treating venous and diabetic ulcers, which often pose challenges in healing. A systematic review and meta-analysis were conducted, evaluating 10 relevant studies involving 633 participants. Findings revealed that HAM treatment significantly accelerated ulcer closure, demonstrating over 90% complete healing compared to standard care. Despite moderate heterogeneity among studies, the results strongly suggested the effectiveness and safety of HAM therapy for venous and diabeticleg ulcers. Further research with larger study cohorts is recommended to bolster the existing evidence supporting HAM in managing these challenging wounds.

Assessment of the feasibility of human amniotic membrane stem cell-derived cardiomyocytes in vitro

Myocardial infarction (MI) is a leading cause of death worldwide, resulting in extensive loss of cardiomyocytes and subsequent heart failure. Inducing cardiac differentiation of stem cells is a potential approach for myocardial regeneration therapy to improve post-MI prognosis. Mesenchymal stem cells (MSCs) have several advantages, including immune privilege and multipotent differentiation potential. This study aimed to explore the feasibility of chemically inducing human amniotic membrane MSCs (hAMSCs) to differentiate into cardiomyocytes in vitro. Human amniotic membrane (AM) samples were obtained from routine cesarean sections at Far Eastern Memorial Hospital. The isolated cells exhibited spindle-shaped morphology and expressed surface antigens CD73, CD90, CD105, and CD44, while lacking expression of CD19, CD11b, CD19, CD45, and HLA-DR. The SSEA-1, SSEA-3, and SSEA-4 markers were also positive, and the cells displayed the ability for tri-lineage differentiation into adipocytes, chondrocytes, and osteoblasts. The expression levels of MLC2v, Nkx2.5, and MyoD were analyzed using qPCR after applying various protocols for chemical induction, including BMP4, ActivinA, 5-azacytidine, CHIR99021, and IWP2 on hAMSCs. The group treated with 5 ng/ml BMP4, 10 ng/ml Activin A, 10 μM 5-azacytidine, 7.5 μM CHIR99021, and 5 μM IWP 2 expressed the highest levels of these genes. Furthermore, immunofluorescence staining demonstrated the expression of α-actinin and Troponin T in this group. In conclusion, this study demonstrated that hAMSCs can be chemically induced to differentiate into cardiomyocyte-like cells in vitro. However, to improve the functionality of the differentiated cells, further investigation of inductive protocols and regimens is needed.

Lyophilised human amniotic membrane allograft modulates cellular activities supportive of wound management.

Lyophilised human amniotic membrane allograft modulates cellular activities supportive of wound management.

Clinical Outcomes and Indications of In-Office Sutureless Dried Gamma Ray-Sterilized Human Amniotic Membrane Transplantation With Bandage Contact Lenses in Various Ocular Surface Disorders.

The purpose of this study was to investigate the efficacy and indications of using dried gamma ray-sterilized amniotic membrane (AM) transplantation with bandage contact lenses for the in-office sutureless treatment of various ocular surface diseases. This study retrospectively included 56 eyes of 52 patients with various ocular surface diseases treated with sutureless dehydrated AM-assisted therapeutic contact lenses. The patients were followed up and assessed 1, 2, and 4 weeks after the treatment, and the therapeutic contact lenses were removed after 1 or 2 weeks. The size of the corneal lesion and the degree of pain experienced by each patient before and after the procedure were measured. Corneal re-epithelization rate and clinical aspects related to re-epithelialization were evaluated. Complete corneal re-epithelialization of the epithelial defect was observed in 40 eyes. The complete re-epithelialization rates of persistent epithelial defect caused by infectious ulcers, neurotrophic ulcers, ulcers due to burn, toxic keratopathy, previous penetrating keratoplasty or other corneal surgeries, and severe dry eye disease were 69.2%, 80.0%, 77.8%, 100%, 75.0%, and 100%, respectively. Five patients with uncontrolled infectious ulcers, neurotrophic ulcer, bullous keratopathy, and ulcers associated with rheumatic disease did not show significant improvement. Patient pain scores significantly decreased after the procedure. Dehydrated AM transplantation with a bandage contact lens is minimally invasive and easily applicable through in-office procedures. This approach is expected to be more actively applied in various ocular surface disorders, not only in severe corneal diseases but also in relatively mild corneal surface diseases.

Sterilization of Human Amniotic Membrane Using an Ozone Hydrodynamic System.

Human amniotic membrane (hAM) is an important biomaterial for Tissue Engineering, due to its great regenerative properties and potential use as a scaffold. The most used procedure to sterilize biomaterials is gamma-irradiation, but this method can affect several properties, causing damage to the structure and reducing the growth factors. The present work evaluated the efficiency of a new method based on ozonated dynamic water for hAM sterilization. HAM fragments were experimentally contaminated with Staphylococcus aureus, Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Clostridium sporogenes (106CFU/mL) and submitted to sterilization process for 5, 10 and 15min. The analyses did not reveal microbial activity after 10min for S. aureus and C. sporogenes and after 15min for E. coli and S. epidermidis. The microbial activity of C. albicans was reduced with the exposure time increase, but the evaluated time was insufficient for complete sterilization. The depyrogenation process was investigated for different ozonation times (15, 20, 25, 30, and 35min) to evaluate the ozone sterilization potential and presented promising results after 35min. The ozone effect on hAM structure was evaluated by histological analysis. A decrease in epithelium average thickness was observed with the exposure time increase. Furthermore, some damage in the epithelium was observed when hAM was exposed for 10 and 15min. It can indicate that ozone, besides being effective in sterilization, could promote the hAM sample's de-epithelization, becoming a possible new method for removing the epithelial layer to use hAM as a scaffold.

Assessment of the proteome profile of decellularized human amniotic membrane and its biocompatibility with umbilical cord-derived mesenchymal stem cells.

Extracellular matrix-based bio-scaffolds are useful for tissue engineering as they retain the unique structural, mechanical, and physiological microenvironment of the tissue thus facilitating cellular attachment and matrix activities. However, considering its potential, a comprehensive understanding of the protein profile remains elusive. Herein, we evaluate the impact of decellularization on the human amniotic membrane (hAM) based on its proteome profile, physicochemical features, as well as the attachment, viability, and proliferation of umbilical cord-derived mesenchymal stem cells (hUC-MSC). Proteome profiles of decellularized hAM (D-hAM) were compared with hAM, and gene ontology (GO) enrichment analysis was performed. Proteomic data revealed that D-hAM retained a total of 249 proteins, predominantly comprised of extracellular matrix proteins including collagens (collagen I, collagen IV, collagen VI, collagen VII, and collagen XII), proteoglycans (biglycan, decorin, lumican, mimecan, and versican), glycoproteins (dermatopontin, fibrinogen, fibrillin, laminin, and vitronectin), and growth factors including transforming growth factor beta (TGF-β) and fibroblast growth factor (FGF) while eliminated most of the intracellular proteins. Scanning electron microscopy was used to analyze the epithelial and basal surfaces of D-hAM. The D-hAM displayed variability in fibril morphology and porosity as compared with hAM, showing loosely packed collagen fibers and prominent large pore areas on the basal side of D-hAM. Both sides of D-hAM supported the growth and proliferation of hUC-MSC. Comparative investigations, however, demonstrated that the basal side of D-hAM displayed higher hUC-MSC proliferation than the epithelial side. These findings highlight the importance of understanding the micro-environmental differences between the two sides of D-hAM while optimizing cell-based therapeutic applications.

Bioengineered Skin for Diabetic Foot Ulcers: A Scoping Review.

Diabetic foot ulcers (DFUs) pose a significant threat to individuals with diabetes mellitus (DM), such as lower limb amputation and severe morbidity. Bioengineered skin substitutes (BSS) are alternatives to traditional interventions for treating DFUs, but their efficacy compared to standard wound care (SWC) or other treatment types, such as allografts, remains unknown. A scoping review of human studies was conducted to identify current approaches in the treatment of DFUs using BSS as compared with other treatment options. Systematic searches in PubMed, Cochrane Library, and Web of Science were conducted to identify comparative studies that enrolled 10 or more patients and evaluated wound healing outcomes (closure, time-to-healing, and area reduction). Database searches isolated articles published from 1 December 2012 to 1 December 2022 and were conducted in accordance with PRISMA-ScR guidelines. The literature search yielded 1312 articles, 24 of which were included for the qualitative analysis. Findings in these studies demonstrated that BSS outperformed SWC in all measured outcomes, suggesting that BSS may be a superior treatment for DFUs. Of the 24 articles, 8 articles compared human amniotic membrane allografts (hAMA) to BSS. Conflicting evidence was observed when comparing BSS and hAMA treatments, highlighting the need for future research.

Silica nanoparticles enhance interfacial self-adherence of a multi-layered extracellular matrix scaffold for vascular tissue regeneration.

Based on the clinical need for grafts for vascular tissue regeneration, our group developed a customizable scaffold derived from the human amniotic membrane. Our approach consists of rolling the decellularized amniotic membrane around a mandrel to form a multilayered tubular scaffold with tunable diameter and wall thickness. Herein, we aimed to investigate if silica nanoparticles (SiNP) could enhance the adhesion of the amnion layers within these rolled grafts. To test this, we assessed the structural integrity and mechanical properties of SiNP-treated scaffolds. Mechanical tests were repeated after six months to evaluate adhesion stability in aqueous environments. Our results showed that the rolled SiNP-treated scaffolds maintained their tubular shape upon hydration, while non-treated scaffolds collapsed. By scanning electron microscopy, SiNP-treated scaffolds presented more densely packed layers than untreated controls. Mechanical analysis showed that SiNP treatment increased the scaffold's tensile strength up to tenfold in relation to non-treated controls and changed the mechanism of failure from interfacial slipping to single-point fracture. The nanoparticles reinforced the scaffolds both at the interface between two distinct layers and within each layer of the extracellular matrix. Finally, SiNP-treated scaffolds significantly increased the suture pullout force in comparison to untreated controls. Our study demonstrated that SiNP prevents the unraveling of a multilayered extracellular matrix graft while improving the scaffolds' overall mechanical properties. In addition to the generation of a robust biomaterial for vascular tissue regeneration, this novel layering technology is a promising strategy for a number of bioengineering applications.

PRETREATED LYOPHILIZED HUMAN AMNIOTIC MEMBRANE GRAFT COVERING FOR RETINAL DETACHMENT WITH POSTERIOR RETINAL BREAKS ABOVE CHORIORETINAL ATROPHY IN PATHOLOGIC MYOPIA.

To compare the surgical results of vitrectomy with untreated or pretreated lyophilized human amniotic membrane grafts covering in treating retinal detachment related to posterior retinal breaks above chorioretinal atrophy in pathologic myopia. Nineteen patients with retinal detachment related to macular hole (MH) located above macular atrophy and/or posterior paravascular retinal breaks (PRBs) located above patchy chorioretinal atrophy in pathologic myopia were included. The eyes of these patients underwent vitrectomy with untreated lyophilized human amniotic membrane covering (n = 10) or perfluorocarbon liquid (PFCL)-assisted pretreated lyophilized human amniotic membrane covering (n = 9; grafts were pretreated in 0.125% indocyanine green and 50% hypertonic glucose solution for 15-20 minutes). The closure of the MH or PRBs, reattachment of the retina, and best-corrected visual acuity were measured postoperatively. Postoperatively, graft dislocation or shift was only found in two eyes (20%) in the untreated group. The closure rate of the MH or PRBs was 80% (8/10) and 100% (9/9) in the untreated group and the pretreated group, respectively. The occurrence rate of excessive gliosis was 40% and 11% in the untreated group and the pretreated group, respectively. In both groups, best-corrected visual acuity was improved and the retinal reattachment rate was 100% at the final visit. Perfluorocarbon liquid-assisted pretreated lyophilized human amniotic membrane graft covering was effective in treating retinal detachment related to MH and/or PRBs situated above macular atrophy or patchy chorioretinal atrophy in pathologic myopia. This technique appeared to reduce graft dislocation or shift, promote the closures of MHs/PRBs, and reduce the occurrence of gliosis.

Silica nanoparticles enhance the cyto- and hemocompatibility of a multilayered extracellular matrix scaffold for vascular tissue regeneration.

The limited availability of autologous vessels for vascular bypass surgeries is a major roadblock to treating severe cardiovascular diseases. Based on this clinical priority, our group has developed a novel engineered vascular graft by rolling human amniotic membranes into multilayered extracellular matrixes (ECM). When treated with silica nanoparticles (SiNP), these rolled scaffolds showed a significant improvement in their structural and mechanical properties, matching those from gold standard autologous grafts. However, it remained to be determined how cells respond to SiNP-treated materials. As a first step toward understanding the biocompatibility of SiNP-dosed biomaterials, we aimed to assess how endothelial cells and blood components interact with SiNP-treated ECM scaffolds. To test this, we used established in vitro assays to study SiNP and SiNP-treated scaffolds' cyto and hemocompatibility. Our results showed that SiNP effects on cells were concentration-dependent with no adverse effects observed up to 10μg/ml of SiNP, with higher concentrations inducing cytotoxic and hemolytic responses. The SiNP also enhanced the scaffold's hydrophobicity state, a feature known to inhibit platelet and immune cell adhesion. Accordingly, SiNP-treated scaffolds were also shown to support endothelial cell growth while preventing platelet and leukocyte adhesion. Our findings suggest that the addition of SiNP to human amniotic membrane extracellular matrixes improves the cyto- and hemocompatibility of rolled scaffolds and highlights this strategy as a robust mechanism to stabilize layered collagen scaffolds for vascular tissue regeneration.

Case Report and Review of the Literature: Human Amniotic Membrane in the Management of Burn Wounds

High-voltage electrical burns often have a deeper extent and cause more damage to surrounding soft tissue than injuries caused by fire burns or scalds. For the treatment of burn injuries there are dressings, dermal analogues, temporary skin substitutes; which favor the re-epithelialization of the wound bed, accelerating the healing process and some of them having specific properties. We present a case of a 41-year-old man who suffered high-voltage electrical burns on the face, neck, thorax, abdomen, upper limbs, and lower limbs. He underwent surgical cleaning at the day of his hospital admission, 7 days later a second surgical cleaning was performed with placement of human amniotic membrane in thoracic and abdominal burns. 10 months after the burn, the patient presents complete recovery of his burn injuries, hyperchromic scars with no evidence of pathological healing. As neurological sequelae, he presented polyneuropathy secondary to electrocution. We conclude that the amniotic membrane is an excellent substitute for temporary skin, useful for promoting the epithelialization of bloody areas from burns and reducing the local inflammatory response. Conclusion: The comparison between robotic-assisted liver surgery and laparoscopic surgery shows that the average medical cost was higher in robotic surgery when compared to laparoscopic liver cancer.

Human keratin matrices promote wound healing by modulating skin cell expression of cytokines and growth factors.

A wide variety of biomaterials has been developed to assist in wound healing, including acellular animal and human-derived protein matrices. However, millions of patients worldwide still suffer from non-healing chronic wounds, demonstrating a need for further innovation in wound care. To address this need, a novel biomaterial, the human keratin matrix (HKM), was developed, characterised, and tested in vitro and in vivo. HKM was found to be degradation-resistant, and a proteomics analysis showed it to be greater than 99% human keratin proteins. PCR revealed adult human epidermal keratinocytes (HEKa) grown in contact with HKM showed increased gene expression of keratinocyte activations markers such as Epidermal Growth Factor (EGF). Additionally, a cytokine microarray demonstrated culture on HKM increased the release of cytokines involved in wound inflammatory modulation by both HEKa cells and adult human dermal fibroblasts (HDFa). Finally, in a murine chronic wound model, full-thickness wounds treated weekly with HKM were smaller through the healing process than those treated with human amniotic membrane (AM), bovine dermis (BD), or porcine decellularized small intestinal submucosa (SIS). HKM-treated wounds also closed significantly faster than AM- and SIS-treated wounds. These data suggest that HKM is an effective novel treatment for chronic wounds.

A biosynthetic alternative to human amniotic membrane for cornea repair—A comparative in‐vitro study

Aims/Purpose: We hypothesize that the biosynthetic Symatix membrane (SM) can replace human amniotic membrane (HAM) in ocular surgical applications. Our aim was to test this hypothesis by comparing the performance of limbal epithelial cells (LECs) cultures on the membranes.Methods: SM was manufactured by the Electrospinning Company Ltd using its proprietary Symatix® Technology platform. SM consists of degradable medical grade poly (lactic‐co‐glycolic acid) polymer fibres and sodium hyaluronan. The suturing and fibrin gluing of SM were tested ex‐vivo by simulation on human corneas. LECs from human limbal tissue were used for testing cellular properties namely cell proliferation (scratch wound and Ki‐67 assay), metabolic activity, limbal epithelial cell marker (cytokeratin‐19 (CK‐19)) tight junction formation (zonula occludens‐1 (ZO‐1)), cell‐ECM interaction (fibronectin (FN)), and basement membrane protein expression (laminin) on SM, plastic, HAM, and freeze‐dried amniotic membrane (FDAM).Results: The surgical handleability of SM was equivalent to HAM. Histological studies demonstrated that epithelial cells on SM had the typical tightly apposed, polygonal, corneal epithelial cell morphology. Rapid wound healing occurred on SM within 3 days; Ki‐67 revealed increased progressive proliferation; and improved metabolic activity was found on SM compared to HAM and FDAM. Immunofluorescence studies showed positive expression of CK‐19, collagen‐1 (Col‐1), laminin, ZO‐1, and FN in SM, plastic, and FDAM compared to negative expression of CK‐19 and ZO‐1 and positive expression of collagen‐1 (Col‐1), laminin and FN in HAM. This could indicate that SM is a better substrate for epithelial cell migration, proliferation, and tight junction formation.Conclusions: This data demonstrates that SM can provide a suitable alternative to HAM for surgical application in sight‐restoring operations.