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Los procedimientos endodónticos regenerativos (REPs) representan una evolución significativa en el campo de la endodoncia, buscando no sólo tratar la infección o lesión presente en el diente, sino tam-bién promover la regeneración de los tejidos denta-rios afectados. El presente caso clínico muestra un incisivo lateral superior izquierdo con apexogénesis incompleta y diagnóstico de absceso alveolar crónico reagudizado en una paciente de 22 años, en el que se aplicó un procedimiento de endodoncia regenerativa (REPs). La estrategia terapéutica elegida se basó en los principios de ingeniería tisular, incorporando la novedosa aplicación de la membrana amniótica hu-mana liofilizada esterilizada como andamio bioactivo intraconducto. Las evaluaciones clínicas, radiográ-ficas y tomográficas a corto, mediano y largo plazo revelaron el éxito de la terapia. La resolución exitosa mostró en los controles a la pieza dentaria asintomá-tica, con una notable remisión de la patología apical, aumento de la longitud radicular y disminución del calibre apical. Se ha podido destacar la eficacia de los REPs, con una exitosa aplicabilidad de la membra-na amniótica como andamio innovador (AU)
Regenerative endodontic procedures (REPs) represent a significant evolution in the field of endodontics, aiming not only to address the infection or injury within the tooth, but also to promote the regeneration of the affected dental tissues. In this clinical case, an upper left lateral incisor with incomplete apexogenesis and diagnosis of acute exacerbation of a chronic periapical lesion in a 22-year-old patient is presented. A regenerative endodontic procedure (REPs) was applied. The chosen therapeutic strategy was based on tissue engineering principles, incorporating the innovative use of sterilized lyophilized human amniotic membrane as an intraconduct bioactive scaffold. Clinical, radiographic, and tomographic assessments at short, medium, and long-term follow-up revealed the success of the therapy. Successful resolution demonstrated an asymptomatic tooth in the follow-up, with a notable resolution of apical pathology, increased root length, and decreased apical caliber. The effectiveness of REPs has been highlighted, demonstrating the successful applicability of amniotic membrane as an innovative scaffold (AU)
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Humanos , Feminino , Adulto , Células-Tronco/fisiologia , Alicerces Teciduais , Argentina , Faculdades de Odontologia , Papila Dentária , Liofilização/métodosRESUMO
Las quemaduras son un problema de interés en salud pública ya que generan un alto índice de morbimortalidad a nivel mundial, las quemaduras térmicas son las más prevalentes y pueden alterar la integridad anatómica, funcional y estética de la piel, aspectos fundamentales para la autoestima del paciente y su capacidad para reintegrarse a la sociedad. Al revisar la literatura sobre el tratamiento de estas afecciones encontramos diversos tratamientos, entre ellos el uso de membrana amniótica humana, la cual ha tenido un impacto importante en el manejo de quemaduras al funcionar como andamio biológico con cualidades regenerativas y antiinflamatorias. El presente artículo tiene como objetivo sintetizar la información actual que describe las aplicaciones de membranas amnióticas humanas en quemaduras, realizamos una revisión exploratoria sistemática de la literatura desde 2010 hasta 2021.
Burns are a problem of interest in public health since they generate a high rate of morbidity and mortality worldwide, thermal burns are the most prevalent and can alter the anatomical, functional and aesthetic integrity of the skin, fundamental aspects for the patient's self-esteem and their ability to reintegrate into society. At review literature about the treatment of these conditions, we find various treatments, including the use of human amniotic membrane, which has had a significant impact on burn management by functioning as a biological scaffold with regenerative and anti-inflammatory qualities. The present article aims to synthesize the current information that describes the applications of human amniotic membranes in burns. We carry out a systematic exploratory review of the literature from 2010 to 2021.
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Purpose: To compare and study the clinical outcome of tarsorrhaphy and amniotic membrane transplant in the healing of persistent corneal epithelial defects in terms of clinical improvement and symptomatic relief. Methods: This was an interventional, prospective study in which a total of 60 patients with persistent epithelial defects (PED's), randomly divided into two groups of 30 patients each who underwent tarsorrhaphy (Group A) or amniotic membrane transplantation (Group B) with a 4-week-follow-up period, were included. The main parameters studied were the size of an epithelial defect, total healing time, pain score, and complications. Results: The study included 60 eyes of 60 patients with PED. The healing time was 9.83 � 6.51 days in Group A (median = 9.50 days, IQR = 1�days) vs. 18.33 � 13.46 days (median = 19.50 days, IQR = 1� days) in Group B. A total of ten eyes (16.7%) did not heal at the end of 4 weeks. Conclusion: There was a significant reduction in the area of epithelial defect at the end of the 1 week and 2 week follow up postoperatively, in both the treatment forms. The mean healing time in patients of Group A was less as compared to that of the patients in Group B.
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Purpose: Our previous study demonstrated the drug reservoir function of human amniotic membrane (HAM) using stable moxifloxacin as a model drug. The purpose of the present study is to evaluate whether HAM can be used as a drug carrier for extended release of extemporaneous preparation of cefazolin. Methods: HAM Buttons (1 Control, 5 Test) were incubated in a freshly prepared (1 ml) sterile topical solution of cefazolin 5% (w/v) for 3 h and 24 h at two different temperatures. The groups were designated as follows: Group IA: Soaking duration 3 h at 4°C; Group IB: Soaking duration 3 h at room temperature; Group IIA: Soaking duration 24 h at 4°C; and Group IIB: Soaking duration 24 h at room temperature. The release kinetics of cefazolin from different groups of drug-laden HAM was studied for a period of 5 days. Samples were assayed for estimation of cefazolin content at different time intervals by High Performance Liquid Chromatography (HPLC) with Photodiode array (PDA) detector. Results: Three-hour cefazolin treatment with HAM at 4°C caused high drug entrapment (24%) compared to room temperature (11%; P < 0.005); however, the release kinetics was not significantly different between Group IA and IB as well as Group IIA and IIB up to the study period. Increase in drug treatment duration did not show increase in entrapment, but caused two-fold (IA Vs IIA) and 1.6-fold (IB Vs IIB) less drug entrapment at 4°C and room temperature, respectively. Conclusion: The results reveal that HAM may be a suitable drug carrier for extended delivery of fortified formulations without compromising stability.
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Objective: Inducing human amniotic membrane mesenchymal stem cells (hAMSCs) to Schwann cells-like cells (SCs-like cells) in vitro, and to evaluate the efficacy of transplantation of hAMSCs and SCs-like cells on nerves regeneration of the rat flaps.
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Abstract Objective Human amniotic membrane (HAM) used as a wound coverage for more than a century. The aim of this study is to evaluate the efficacy of amniotic membrane on wound healing and reduce post-operative complication. Study design Randomized clinical trial study. Place and duration of study Surgery Department, Shahid Faghihi Hospital, Shiraz, in the period of between Sep. 2014 and Nov. 2015. Methodology 73 patients with anal fistula were divided into two groups. The patients suffered from simple perianal fistula (low type) without any past medical history. Fistulotomy were performed for all of them and in interventional group HAM were applied as biologic dressing. Their wound healing improvement was evaluated post-operative in two groups. Results From 73 patients participated in the study, 36 patients were in control group and 37 patients were in intervention group. According to the analysis of images taken from the wound, the rate of wound healing was 67.39% in intervention group and 54.51% in control group (p < 0.001). Discharge, pain, itching and stool incontinency was lower in intervention group. Analysis of pathology samples taken from the wound showed no differences between two groups. Conclusion HAM application could lead to improvement of wound healing and reduced post-operative complications. In conclusion, HAM may act as a biologic dressing in the patients with anal fistula.
Resumo Objetivo Membrana amniótica humana (MAH) tem sido usada para cobrir feridas por mais de um século. O objetivo deste estudo é avaliar a eficácia da membrana amniótica na cicatrização de feridas e reduzir complicações pós-operatórias. Desenho do estudo Ensaio clínico randomizado. Local e duração do estudo Departamento de Cirurgia, Shahid Faghihi Hospital, Shiraz, Irã, entre setembro de 2014 a novembro de 2015. Método 73 pacientes com fístula anal foram divididos em dois grupos. Os pacientes sofriam de fístula perianal simples (tipo baixo) sem histórico médico prévio. A fistulotomia foi realizada em todos eles e no grupo intervenção, MAH foi aplicada como curativo biológico. A melhora da cicatrização foi avaliada no período pós-operatório em dois grupos. Resultados De 73 pacientes que participaram do estudo, 36 pacientes eram do grupo controle e 37 pacientes do grupo intervenção. De acordo com a análise das imagens da ferida, a taxa de cicatrização foi 67,39% no grupo intervenção e 54,51% no grupo controle (p < 0,001). Secreção, dor, prurido e incontinência fecal foi menor no grupo intervenção. A análise das amostras patológicas retiradas da ferida não mostrou diferenças entre os dois grupos. Conclusão A aplicação de MAH pode levar à melhoria da cicatrização de feridas e reduzir as complicações pós-operatórias. Em conclusão, a MAH pode atuar como um curativo biológico nos pacientes com fístula anal.
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Humanos , Masculino , Feminino , Fístula Retal/cirurgia , Âmnio/lesões , Complicações Pós-Operatórias/cirurgia , Cicatrização/fisiologia , Curativos BiológicosRESUMO
Combination between tissue engineering and other fields has brought an innovation in the area of regenerative medicine which ultimate aims are to repair, improve, and produce a good tissue construct. The availability of many types of scaffold, both synthetically and naturally have developed into many outstanding end products that have achieved the general objective in tissue engineering. Interestingly, most of this scaffold emulates extracellular matrix (ECM) characteristics. Therefore, ECM component sparks an interest to be explored and manipulated. The ECM featured in human amniotic membrane (HAM) provides a suitable niche for the cells to adhere, grow, proliferate, migrate and differentiate, and could possibly contribute to the production of angiogenic micro-environment indirectly. Previously, HAM scaffold has been widely used to accelerate wound healing, treat bone related and ocular diseases, and involved in cardiovascular repair. Also, it has been used in the angiogenicity study, but with a different technical approach. In addition, both side of HAM could be used in cellularised and decellularised conditions depending on the objectives of a particular research. Therefore, it is of paramount importance to investigate the behavior of ECM components especially on the stromal side of HAM and further explore the angiogenic potential exhibited by this scaffold.
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Humanos , Âmnio , Matriz Extracelular , Medicina Regenerativa , Engenharia Tecidual , CicatrizaçãoRESUMO
The aim of this study was to determine the efficiency of different human amniotic membrane (HAM) processing methods on the concentration, purity and integrity of RNA. Two different techniques (Technique 1 and Technique 2) were employed for the processing of HAM, which differed in terms of washing solution, sample storage conditions and processing time. Based on preservation of HAM, three groups were formed under each technique. In Technique 1, the groups were fresh frozen 1 (F1), glycerol preserved (GP) and gamma irradiated glycerol preserved (IGP); where else in Technique 2, the groups were fresh frozen 2 (F2), 50% glycerol/Dulbecco’s modified Eagle medium (DMEM) cryopreserved HAM diluted with phosphate buffered saline (GB) and 50% glycerol/DMEM cryopreserved HAM diluted with diethylprocarbonate water (GD). Total RNA was extracted from the samples and their concentration, purity and integrity were examined. The F2 sample of which there was no pre-washing step and involved direct sample storage at -80ºC, shorter processing time and chilled processing conditions had yielded better quality of RNA compared to the others.
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OBJECTIVE: Epidural fibrosis and adhesion are the main reasons for post-laminectomy sustained pain and functional disability. In this study, the authors investigate the effect of irradiated freeze-dried human amniotic membrane on reducing epidural adhesion after laminectomy on a rat model. METHODS: A total of 20 rats were divided into two groups. The group A did not receive human amniotic membrane implantation after laminectomy and group B underwent human amniotic membrane implantation after laminectomy. Gross and microscopic findings were evaluated and compared at postoperative 1, 3 and 8 weeks. RESULTS: The amount of scar tissue and tenacity were reduced grossly in group of rats with human amniotic membrane implantation (group B). On a microscopic evaluation, there were less inflammatory cell infiltration and fibroblast proliferation in group B. CONCLUSION: This experimental study shows that implantation of irradiated freeze-dried human amniotic membrane reduce epidural fibrosis and adhesion after spinal laminectomy in a rat model.
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Animais , Humanos , Ratos , Âmnio , Cicatriz , Síndrome Pós-Laminectomia , Fibroblastos , Fibrose , LaminectomiaRESUMO
A case of premalignant lesion- leukoplakia of the left buccal mucosa, was excised and defect was reconstructed with human amniotic membrane graft. We evaluated the effectiveness of HAM as a grafting material for the reconstruction of oral mucosal defect after surgical excision of leukoplakia. After 4 weeks of grafting procedure, mucosal defect was restored successfully without any complications.
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Background & objectives: Ex vivo expansion of the limbal epithelial cells activates the nerve growth factor (NGF) mediated downstream signal transduction pathway. It is not clear as to which factors control the stemness of the corneal limbal stem cells, i.e., the maintenance of stem cell properties. It is likely that various signaling pathways are involved, including Notch, Wnt and NGF signaling, etc. In the present study, we investigated the activation of phosphoinositide-3-kinase (PI3K) pathway on cells cultured over the chitosan matrix, chitosan silver matrix, chitosan gold matrix, intact and denuded human amniotic membrane (HAM). Methods: Human limbal biopsies obtained from the cadaveric donor eyes were used in this study. The cells cultured over different substrate and observed for the activation of the downstream signaling molecules of PI3K/Akt/FKHRL1 pathway. Western blotting was done to prove the results. Results: The cells cultured over the intact HAM showed the activation of the downstream signaling molecules of PI3K/Akt/JNK pathway compared to the cells grown over other substrates. On inhibition of the PI3K activity there was absence of phosphorylation of downstream effectors in the limbal epithelial cells from the explant culture over the intact HAM. Interpretation & conclusions: The ex vivo expansion of human limbal epithelial progenitor cells on intact HAM was mediated by PI3K/Akt/FKHRL1 pathway, which is known to govern cell survival, and the mitogen-activated protein kinase (MAPK) pathway, known to control the cell mitosis.
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PURPOSE: To establish the immortalized human corneal endothelial cell line (IHCEn) by transducing human papilloma virus (HPV) 16 E6/E7 oncogenes, and to identify their characteristics when cultivated on a lyophilized human amniotic membrane (LAM). METHODS: Primary human corneal endothelial cells (PHCEn) were infected using a retroviral vector with HPV 16 E6/E7, and transformed cells were clonally selected by G418. Growth properties and characteristics of IHCEn were compared with PHCEn by cell counting and RT-PCR of VDAC3, SLC4A4, CLCN3, FGF-1, Col IV, and Na+/K+ ATPase. IHCEn were cultured on LAM. Messenger RNA expressions of VDAC3, CLCN3, and Na+/K+ ATPase, and protein expressions of Na+/K+ ATPase and Col IV in IHCEn cultivated on LAM were investigated by RT-PCR, immunofluorescence, and immunohistochemical staining, respectively. RESULTS: Successful immortalization was confirmed by stable expression of HPV 16 E6/E7 mRNA by RT-PCR, and IHCEn exhibited typical corneal endothelial morphology. Doubling time of IHCEn was 30.15+/-10.96 hrs. Both IHCEn and PHCEn expressed VDAC3, CLCN3, SLC4A4, FGF-1, Col IV, and Na+/K+ ATPase. IHCEn cultivated on LAM showed stronger expression of VDAC3, CLCN4, and Na+/K+ ATPase mRNA than on plastic culture dish. Immunohistochemical staining and immunofluorescence revealed the positive expression of Na+/K+ ATPase and Col IV. CONCLUSIONS: IHCEn were successfully established, and LAM is a good substrate for the culture of human corneal endothelial cells.
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Humanos , Transfecção , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas Repressoras/genética , RNA Mensageiro/genética , Proteínas Tirosina Quinases , Proteínas Oncogênicas Virais/genética , Imuno-Histoquímica , Regulação Viral da Expressão Gênica , Liofilização , Endotélio Corneano/citologia , Linhagem Celular Transformada , Contagem de Células , ÂmnioRESUMO
Objective: To evaluate the microbial safety of human amniotic membrane(HAM) in preparation and preservation,and to establish effective methods of microbe control for HAM bank.Methods: Human amnions were collected from elective caesarean sections in 20 healthy women,and divided into groups based on different sterilization procedures and preservation methods.Bacteria,fungi and mycoplasmas were detected at 24h,1m,3m and 12m after preservation.Other amnions collected in eutocia from 5 healthy women were detected bacteria,fungi,lactobacilli and mycoplasmas.The series of serological determination for the parturients were followed up in 6 months.Results: Two HAM samples only soaked in NS were detected to be contaminated with E.coli and/or S.epidermidis,respectively from two caesarean sections.There is no microbe detected in all of the amnions from caesarean sections which were soaked in the antibiotics solution.One HAM sample from eutocia was yet detected to have lactobacilli after soak.The series of serological determination were all negative.Conclusion: During one year storage,HAM only from caesarean sections in healthy women,can successfully avoid contamination if soaked in the antibiotics solution then prepared and preserved with aseptic technique.