Effects of Preservation Methods in the Composition of the Placental and Reflected Regions of the Human Amniotic Membrane

2021 ◽  
pp. 1-11
Author(s):  
Jéssica Tereza Guedes de Oliveira Moraes ◽  
Maíra Maftoum Costa ◽  
Paula Cristina Santos Alves ◽  
Luciana Barros Sant’Anna
Keyword(s):  
Regenerative Medicine ◽  
Amniotic Membrane ◽  
Structural Integrity ◽  
Storage Period ◽  
Soluble Proteins ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
Glycerol Solution ◽  
Preservation Methods ◽  
Preservation Method

The human amniotic membrane (AM) is emerging as an interesting biomaterial for regenerative medicine due to its biological and mechanical proprieties. The beneficial effects of the AM are probably related to its bioactive factors produced by local cells and stored in the stromal matrix. However, the search for a preservation method capable of preserving AM properties remains a challenge. The aim of this study was to evaluate important features of 2 anatomical regions of the human AM (reflected and placental amnion) after different preservation methods. For this purpose, human placentas were harvested and processed for AM isolation and storage at 2 different conditions: room temperature for 18 h in DMEM (fresh AM) and −80°C in DMEM/glycerol solution for 30 days (cryopreserved AM). After the storage period, the structural integrity of the membrane was assessed by histological and Picrosirius polarization analysis, cellular viability analysis was performed using the MTT assay, and the soluble proteins were quantified with the Qubit Protein Assay Kit. Both preservation protocols reduced the cell viability, mainly in the placental amnion region of the AM, but preserved the morphology of epithelial and stromal layers, as well as the organization and distribution of collagen fibers. There was a reduction in soluble proteins only in fresh AM. Importantly, the cryopreserved AM group presented the same concentration as the control group. In conclusion, the cryopreservation using DMEM/glycerol was ideal for preserving the structural integrity and soluble protein content, indicating the feasibility of this method in preserving AM for its use in regenerative medicine.

scholarly journals Functional Evaluation of Sciatic Nerve Anastomosis Wraped by Freeze Dried Human Amniotic Membrane in Sprague Dawley Rat

2020 ◽  
Vol 2 (1) ◽  
pp. 01-05
Author(s):  
Eko Perdana Putra ◽  
Pamudji Utomo ◽  
Mujaddid Idulhaq
Keyword(s):  
Sciatic Nerve ◽  
Amniotic Membrane ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
Functional Evaluation ◽  
Sprague Dawley ◽  
Group I ◽  
Freeze Dried ◽  
Significant Difference ◽  
Sciatic Functional Index

Background. Peripheral nerve injury (PNI) is a common medical condition. The defected nerve, if not repaired as early as possible, can cause long-term denervation and neurotrophy failure for the target organ. This leads to a series of denervation manifestations, such as muscle atrophy, loss of sensory function, etc. and ultimately, these manifestations seriously affect the patient’s sensorimotor function.1,2  Amniotic membranes have been widely used in ophthalmology and skin injury repair because of their anti-inflammatory properties. In this study, we measured therapeutic efficacy and determined if amniotic membranes could be used for sciatic nerve repair.Material and methods. A post test only control group design has been done  in 10 healthy Sprague Dawley rats. In all rats, a unilateral right side sciatic nerve transection was performed and reanastomosed by different methods: Group I (control group): included 5 rats, the anastomosis was done by epineural microsutures using 8/0 nylon. Group II: included 5 rats, the anastomosis was done by epineural microsutures using 8/0 nylon and  then wraped by freeze dried human amniotic membrane. Functional evaluation of nerve recovery was done over 3 weeks postoperatively using walking tract analysis and calculate using Sciatic Functional Index.Result. Functional results showed that there was no significant difference of the sciatic functional index (SFI) between group I and group II.Conclusion. We can conclude that during 3 weeks functional evaluation, there is no significant difference between control group and  experimental group that achieved freeze dried human amniotic membrane.Keywords: sciatic nerve injury, freeze dried human amniotic membrane, walking tract analysis, sciatic functional index.

Human Amniotic Membrane as a Biological Source for Regenerative Medicine

2016 ◽  
pp. 81-105 ◽  
Author(s):  
Mazaher Gholipourmalekabadi ◽  
Narendra Pal Singh Chauhan ◽  
Behrouz Farhadihosseinabad ◽  
Ali Samadikuchaksaraei
Keyword(s):  
Regenerative Medicine ◽  
Amniotic Membrane ◽  
Human Amniotic Membrane ◽  
Biological Source

scholarly journals Healing Effects of Human Amniotic Membrane and Burned Wool on the Second-degree Burn in Rats

2020 ◽  
Vol 9 ◽  
pp. 1759
Author(s):  
Nader Tanideh ◽  
Farideh Keshavarzi ◽  
Aida Hemat Zadeh ◽  
Sajad Daneshi ◽  
Omid Koohi-Hosseinabadi ◽  
...  
Keyword(s):  
Wound Healing ◽  
Amniotic Membrane ◽  
Granulation Tissue ◽  
Male Rats ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
Tissue Formation ◽  
Granulation Tissue Formation ◽  
Degree Burn ◽  
Second Degree

ackground: This study aimed to compare sheep burnt wool and human amniotic membrane (AM) on second-degree burn wound healing in rats. Materials and Methods: Seventy-two adult male rats of Sprague Dawley underwent general anesthesia, and a deep second-degree burn was created on their skin by a hot iron plate. Afterward, human AM, silver sulfadiazine ointment (SSD), and sheep burned wool were used on wound area for burn treatment. On days 7, 14, and 21 of the experiment, the rats were sacrificed, and histopathological assessments were done. Results: Human AM, in comparison with other groups, significantly (P<0.05) showed better improvement in all pathologic variables. Burned wool showed significant improvement compared to the control group on day 7 in the angiogenesis, on day 14 in granulation tissue formation and epithelial formation, and on day 21 in new epithelial formation (P<0.05). Burned wool compared with SSD ointment in granulation tissue formation improved significantly (P<0.05) on days 7 and 14. Also, SSD ointment in comparison with the control group significantly improved (P<0.05) granulation tissue formation and macrophage on day 7. Conclusion: Human AM has a significant effect on the treatment of second-degree burn. Burned wool has a better effect on wound healing than SSD ointment and negative control group without treatment in terms of granulation tissue and epithelium formation. [GMJ.2020;9:e1759]

scholarly journals The use of amniotic membrane in the repair of duodenal wounds in Wistar rats

2010 ◽  
Vol 25 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Luciano Rodrigues Schimidt ◽  
Edson José Cardoso ◽  
Rogério Rodrigues Schimidt ◽  
Luiz Augusto Back ◽  
Maria Beatriz Schiazawa ◽  
...  
Keyword(s):  
Amniotic Membrane ◽  
Wistar Rats ◽  
Muscle Layer ◽  
Wall Structure ◽  
Duodenal Wall ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
New Material ◽  
Group A ◽  
Group B

Purpose: In the search of a new material to repair duodenal wounds, a trial was conducted to assess the behavior of human amniotic membrane in the repair of the duodenal wall in rats. METHODS: Fifty Wistar rats weighing between 250 and 350g, male, were submitted to duodenotomy and randomly distributed into two groups. Group A (n=8) had no treatment and was used as the control group. In Group B (n=42) the duodenal wound was treated with a patch of human amniotic membrane. RESULTS: All animals in Group A died. In Group B no changes were observed with regards to death or the formation of duodenal fistula. All animals presented peritoneal adherences in the region on the duodenal wall repair and intestinal obstruction was observed in two animals. Healing of the duodenal wall in the region of the patch took place progressively as the post-operatory period increased, with regeneration of the mucosa and of the smooth muscle layer. CONCLUSION: From the clinical standpoint, the amniotic membrane proved to be a biological tissue which served as a temporary seal and allowed the wound to heal by second-intention, with re-establishment of the duodenal wall structure.

scholarly journals Human amniotic membrane as an alternative source of stem cells for regenerative medicine

2011 ◽  
Vol 81 (3) ◽  
pp. 162-171 ◽  
Author(s):  
Silvia Díaz-Prado ◽  
Emma Muiños-López ◽  
Tamara Hermida-Gómez ◽  
Claudia Cicione ◽  
M. Esther Rendal-Vázquez ◽  
...  
Keyword(s):  
Stem Cells ◽  
Regenerative Medicine ◽  
Amniotic Membrane ◽  
Human Amniotic Membrane ◽  
Alternative Source

scholarly journals Bone-Marrow Stem Cells and Acellular Human Amniotic Membrane in a Rat Model of Heart Failure

Life ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 958
Author(s):  
Gustavo Gavazzoni Blume ◽  
Paulo André Bispo Machado-Junior ◽  
Rossana Baggio Simeoni ◽  
Giovana Paludo Bertinato ◽  
Murilo Sgarbossa Tonial ◽  
...  
Keyword(s):  
Heart Failure ◽  
Stem Cells ◽  
Bone Marrow ◽  
Amniotic Membrane ◽  
Rat Model ◽  
Connexin 43 ◽  
Left Ventricular ◽  
Bone Marrow Stem Cells ◽  
Control Group ◽  
Human Amniotic Membrane

Myocardial infarction (MI) remains the leading cause of cardiovascular death worldwide and a major cause of heart failure. Recent studies have suggested that cell-based therapies with bone marrow stem cells (BMSC) and human amniotic membrane (hAM) would recover the ventricular function after MI; however, the mechanisms underlying these effects are still controversial. Herein, we aimed to compare the effects of BMSC and hAM in a rat model of heart failure. MI was induced through coronary occlusion, and animals with an ejection fraction (EF) < 50% were included and randomized into three groups: control, BMSC, and hAM. The BMSC and hAM groups were implanted on the anterior ventricular wall seven days after MI, and a new echocardiographic analysis was performed on the 30th day, followed by euthanasia. The echocardiographic results after 30 days showed significant improvements on EF and left-ventricular end-sistolic and end-diastolic volumes in both BMSC and hAM groups, without significant benefits in the control group. New blood vessels, desmine-positive cells and connexin-43 expression were also elevated in both BMSC and hAM groups. These results suggest a recovery of global cardiac function with the therapeutic use of both BMSC and hAM, associated with angiogenesis and cardiomyocyte regeneration after 30 days.

scholarly journals Using Dry Human Amniotic Membrane in Secondary Intention Wound Healing After Urological Cancer Surgery: The First Randomized Clinical Trial in Iran

2021 ◽  
Vol 14 (5) ◽  
Author(s):  
Ramin Hamidi ◽  
Ebrahim Hazrati ◽  
Davood Abbasi Azar ◽  
Farzad Allameh ◽  
Babak Javanmard ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Wound Healing ◽  
Randomized Clinical Trial ◽  
Amniotic Membrane ◽  
Cancer Surgery ◽  
Control Group ◽  
Case Group ◽  
Human Amniotic Membrane ◽  
Urological Cancer ◽  
Secondary Intention

Background: Wound healing is a complex process and in some patients, it is a long process. Due to the presence of various underlying diseases in patients with cancer, this process is delayed, and as a result, the rate of complications increases. Therefore, some materials are needed to accelerate wound healing. Nowadays efficacy of dry human amniotic membrane in burn wounds and chronic wounds is proven; however, no studies have been performed on the effect of this tissue in the treatment of surgical wounds. Objectives: This study aimed at evaluating the efficacy of dry human amniotic membrane in the secondary repair of urological cancer surgery wounds. Methods: In this randomized clinical trial from January 2018 to June 2020, 40 patients who underwent urological cancer surgery and secondary intention wound healing were selected and randomly divided into 2 groups of 20 patients. In the case group, the dry human amniotic membrane was used for dressing wounds, and in the control group, normal dressing without an amniotic membrane was performed. Complications such as infection, sepsis, and reoperation, as well as the duration of wound healing and hospitalization of patients at intervals of 1 week, 1, and 3 months, were compared between the 2 groups. Results: The 2 groups matched in terms of gender, age, and wound location. The mean duration of hospitalization and wound healing was significantly lower in the case group (P < 0.05). The rate of complications was higher in the control group but was not significant. Conclusions: Dry human amniotic membranes due to the accelerated wound healing process and fewer complications could be a good choice for secondary healing of wounds after urological cancer surgery.

scholarly journals Amniotic membrane applied to burns healing: Pre-clinical study

2021 ◽  
Vol 10 (4) ◽  
pp. 38110414286
Author(s):  
Fernanda Cláudia Miranda Amorim ◽  
Emilia Ângela Loschiavo Arisawa ◽  
Luciana Barros Sant’Anna ◽  
Khetyma Moreira Fonseca ◽  
Davidson Ribeiro Costa ◽  
...  
Keyword(s):  
Amniotic Membrane ◽  
Healing Process ◽  
Inflammatory Cells ◽  
Repair Process ◽  
Treated Group ◽  
Preclinical Study ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
Significance Level ◽  
Kolmogorov Smirnov

This preclinical study aimed to evaluate the tissue repair process of burns treated with human amniotic membrane (hAM) patches in rats. Twenty-four rats were subjected to superficial burns of partial thickness, and randomly allocated into two groups: Control and Treated Group, subdivided into two experimental periods of 7th and 14th days. The lesions were evaluated by digitalized images (macroscopy) and by the analysis of histological sections stained in H&E to quantify the number of inflammatory cells and fibroblasts present in the different experimental times (histomorphometry). The histomorphometric analyses were performed blindly. Statistical analysis employed Kolmogorov-Smirnov and Mann Whitney tests, with 95% confidence interval at 5% significance level (p <0.05). Macroscopically, the lesions of Treated group presented a crust formation before Control Group, and there were no signs of infection in both groups. Microscopically, the qualitative analysis showed a faster evolution in the healing process of the Treated groups compared to the Control, with reduction of the inflammatory infiltrate, intense fibroblasts proliferation and better organization of the collagen fibers. The quantitative analysis showed statistically significant results regarding the reduction of inflammatory cells (p<0.0001) at 7th and 14 th day and increased proliferation of fibroblasts at 14th day (p<0.0001) in lesions treated with hAM compared to Control group. The results of this preclinical study demonstrated that the application of hAM patches reduces the inflammatory process and accelerates the onset of the proliferative phase in burn injuries.

scholarly journals Human amniotic membrane grafts for retinal breaks in diabetic tractional retinal detachment and combined tractional and rhegmatogenous retinal detachment

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yen-Chih Chen ◽  
San-Ni Chen
Keyword(s):  
Retinal Detachment ◽  
Tissue Regeneration ◽  
Amniotic Membrane ◽  
Rhegmatogenous Retinal Detachment ◽  
Case Series ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
Ellipsoid Zone ◽  
Retinal Breaks ◽  
Additional Surgery

AbstractTo describe the surgical outcomes of using human amniotic membrane (hAM) grafts in the management of retinal breaks in diabetic tractional detachment (TRD) and combined tractional and rhegmatogenous retinal detachment (CTRRD). A retrospective case series of 10 eyes with TRD or CTRRD receiving pars plana vitrectomy with hAM grafts implantation, compared with 13 controls receiving the same surgery without hAM grafts. Best-corrected visual acuity (BCVA) and re-detachment rate were compared between two groups. Postoperatively, all eyes in the hAM group had retina attachment without recurrence, while 9 eyes in the control group had retina re-detachment and required additional surgery (0% vs 69.2%, p = 0.003). The BCVA significantly improved in the hAM group (from 1.96 ± 0.95 to 1.44 ± 0.77 in log MAR, p = 0.03), but not improved in control group (p = 0.20). Postoperative optical coherence tomography of the eyes receiving hAM grafts demonstrated glial tissue regeneration and restoration of ellipsoid zone. In diabetic TRD or CTRRD, hAM grafts could be an effective method, with promising outcome. Compared to standard surgery, it could result in higher retina reattachment rate and significant visual improvement. Moreover, it may offer the adjunctive benefit in tissue regeneration and fasten ellipsoid zone restoration.

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