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1.
Acta Cir Bras ; 35(1): e202000101, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32159587

RESUMO

Solid organ transplantation is a very complex process, in which the storage of the graft in a preservation solution is mandatory in order to extend ischemic times and contain further damage. The condition in which the organ is transplanted is critical for the outcome of the organ recipient. The recent emergence of generic versions of organ preservation solutions (solutions with the same composition and under the same legislation as the original versions, but with different brands) compelled us to study whether the standards are maintained when comparing the original and its generic counterpart. Along these lines, we discuss and comment on some aspects concerning this issue of general interest in the organ transplantation field.


Assuntos
Glutationa/química , Soluções para Preservação de Órgãos/química , Transplante de Órgãos/métodos , Cálcio/análise , Humanos , Soluções para Preservação de Órgãos/normas , Temperatura Ambiente , Fatores de Tempo
2.
Eur Arch Paediatr Dent ; 21(1): 53-59, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31104259

RESUMO

AIM: The purpose of this study was to investigate the effect of different storage media on viability and proliferation capacity of periodontal ligament cells. METHODS: Plates with periodontal ligament fibroblasts (PDLF) cells were incubated in skimmed and whole milk, recently prepared Hank's balanced salt solution (HBSS), Save-A-Tooth system's, coconut water, propylene glycol with 20% propolis, egg white, tap water (negative control) at 5 °C and 20 °C, for 24 h. In one of the plates of each temperature, cell viability was determined by MTT assay. In the remaining plates, the wells were filled and incubated with Minimum Essential Medium (MEM) at 37 °C for 24, 48, 72, 96, and 120 h. The proliferation capacity of PDFL cells was also evaluated by MTT assay. Data were statistically analyzed by Kruskal-Wallis, Scheffé and Mann-Whitney tests (α = 5%). RESULTS: At 5 °C, milk maintained more viable cells than other storage media immediately after exposure (0 h) and allowed greater proliferation capacity. At 20 °C, milk and HBSS had similar and allowed similar proliferation ability at 24 and 48 h. From 72 h onwards, capacity to maintain cell viability the proliferation rate of cells incubated in HBSS was superior than milk. At both temperature and experiments, Save-A-Tooth system was similar to tap water. CONCLUSION: Milk and HBSS were more effective in maintaining cellular viability and proliferation capacity than any other storage media. At 5 °C, the most viable alternative was milk. At 20 °C, HBSS had better results.


Assuntos
Soluções para Preservação de Órgãos , Avulsão Dentária , Animais , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Meios de Cultura , Humanos , Soluções Isotônicas , Leite , Ligamento Periodontal
3.
Dent Traumatol ; 36(1): 3-18, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31328384

RESUMO

BACKGROUND/AIMS: Dental avulsion is defined as the complete displacement of a tooth from its socket owing to trauma. The treatment outcome depends on storage of the avulsed teeth in media capable of maintaining the viability of periodontal ligament cells, when immediate replantation is not possible. To maintain the viability of periodontal ligament cells, plants can be used as a storage medium because of their pharmacological and phytotherapic properties. The aim of this study was to evaluate the effect of plants on the tissue repair following tooth replantation. METHODS: This systematic review was conducted according to the PRISMA guidelines and included articles collected in the Cochrane, LILACS, PubMed, Science Direct, Scopus and Web of Science databases, plus articles found in the grey literature. The articles were screened for partial reading using the Endnote and Rayyan platform. The methodology of studies was evaluated by using the OHAT and GRADE. RESULTS: In the initial search, 2361 articles were obtained, only 51 articles were submitted to complete reading, and 35 articles were selected for the qualitative analysis. The evaluated plants had a potential effect on cell viability and proliferation. The articles evaluated mainly the action of plants on cells of the periodontal ligament. Propolis, coconut water and Aloe vera were the most common storage medium. CONCLUSION: The methodological limitations persist, and the evaluation of the pharmacological potential of plants on dental tissues still requires more research.


Assuntos
Aloe , Cocos , Soluções para Preservação de Órgãos , Própole , Avulsão Dentária , Humanos , Ligamento Periodontal , Reimplante Dentário
4.
Dent Traumatol ; 36(1): 58-68, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31050380

RESUMO

BACKGROUND/AIM: Various types of storage media have been investigated to preserve avulsed teeth. However, the efficacies of storage media mainly focus on the aspect of cell viability. The aim of this study was to evaluate and compare the gene expression profiles of human periodontal ligament cells preserved in Hank's balanced salt solution (HBSS) and milk over different storage durations. MATERIAL AND METHODS: Human periodontal ligament cells were cultured and preserved in HBSS and milk for 3 and 6 hours. Next, total RNA was isolated. QuantSeq 3' mRNA-Sequencing was used to examine differences in gene expression in HBSS- and milk-grown periodontal ligament cells. Bioinformatics analysis was also performed to predict the function of the differentially expressed genes. RESULTS: The number of differentially expressed genes shared among all groups was 101. In gene set enrichment analysis, the shared differentially expressed genes in HBSS and milk were associated with the TNF-α signaling pathway (P = 1.07E-7 ). Seven hallmark gene sets were also identified in HBSS. Moreover, hallmark gene sets associated with hypoxia (P = 7.26E-5 ) and apoptosis (P = 4.06E-4 ) were identified in HBSS. In milk, 10 hallmark gene sets along with gene sets for inflammatory response (P = 6.87E-3 ) were identified. CONCLUSIONS: Compared to those in milk, genes in HBSS were differentially expressed with increasing storage duration, suggesting that diverse and different gene expression may be involved in HBSS and milk. However, a more detailed functional analysis of these differentially expressed genes in storage solutions should be performed in the future.


Assuntos
Leite , Soluções para Preservação de Órgãos , Ligamento Periodontal , Avulsão Dentária , Transcriptoma , Animais , Sobrevivência Celular , Células Cultivadas , Humanos , Soluções Isotônicas , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo
5.
J Endod ; 46(1): 74-80, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31843129

RESUMO

INTRODUCTION: Histidine-tryptophan-ketoglutarate (HTK) is a preservation solution used for organ transplantation. The physiological pH and osmolality of this solution are known to facilitate cell proliferation and cell membrane stabilization. The purpose of the present study was to investigate the efficacy of several concentrations of HTK solution as a storage medium for avulsed teeth. METHODS: Cultured human periodontal ligament cells were stored in different concentrations of HTK solutions. After 1, 3, 6, 12, 24, 48, and 72 hours, cell viability was assessed using the Cell-Counting Kit-8 (Dojindo Molecular Technologies, Kumamoto, Japan) and LIVE/DEAD (Invitrogen, Carlsbad, CA) assay. Cell response of the most effective concentrations of HTK solution were further analyzed by gene expression profiling, and their cell viability was compared with other storage media. RESULTS: The highest cell viability was observed in 50% HTK solution in various concentrations of HTK solution (P < .05). In periodontal ligament cells stored in 50% HTK solution for 3 hours, the expression of genes related to angiogenesis, the inflammatory response, and cell proliferation was increased compared with the control. Compared with other storage media, the highest cell viability was observed in 50% HTK solution. CONCLUSIONS: Our study suggests that 50% HTK solution containing cell culture medium represents a suitable storage medium for avulsed teeth.


Assuntos
Soluções para Preservação de Órgãos , Avulsão Dentária , Glucose , Glutationa , Humanos , Insulina , Manitol , Preservação de Órgãos , Cloreto de Potássio , Procaína
6.
Pediatr Dent ; 41(6): 485-488, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31882036

RESUMO

Purpose: The purpose of this study was to compare the effects of virgin olive oil (VOO), soybean oil (SO), and Hank's Balanced Salt Solution (HBSS) on the vitality of periodontal ligament (PDL) cells of simulated avulsed teeth. Methods: Forty freshly extracted teeth were randomly divided into three experimental groups (n equals 10), one positive control group (n equals five), and one negative control group (n equals five). The experimental teeth were air-dried for 30 minutes and then soaked in one of the three storage solutions: HBSS, VOO, or SO. To quantify the number of viable cells, a collagenase-dispase assay was used. The viable PDL cells were determined via 0.4% Trypan blue staining. Data were statistically analyzed using the Kruskal-Wallis H test and Mann-Whitney U test with a significance level of 0.05. Results: The number of viable cells was significantly higher after storage in SO than in HBSS (P=0.004). There was no significant difference between SO and VOO in terms of PDL cell viability. Conclusion: Vegetable oils can be promising storage solutions for maintaining the periodontal ligament cell viability of avulsed teeth.


Assuntos
Olea , Soluções para Preservação de Órgãos , Avulsão Dentária , Sobrevivência Celular , Humanos , Soluções Isotônicas , Leite , Azeite de Oliva , Ligamento Periodontal , Óleo de Soja
7.
J Cardiothorac Surg ; 14(1): 174, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31615560

RESUMO

BACKGROUND: Vein graft disease (VGD) impairs graft patency rates and long-term outcomes after coronary artery bypass grafting (CABG). DuraGraft is a novel endothelial-damage inhibitor developed to efficiently protect the structural and functional integrity of the vascular endothelium. The DuraGraft registry will evaluate the long-term clinical outcomes of DuraGraft in patients undergoing CABG procedures. METHODS: This ongoing multicentre, prospective observational registry will enrol 3000 patients undergoing an isolated CABG procedure or a combined procedure (ie, CABG plus valve surgery or other surgery) with at least one saphenous vein grafts or one free arterial graft (ie, radial artery or mammary artery). If a patient is enrolled, all free grafts (SVG and arterial will be treated with DuraGraft. Data on baseline, clinical, and angiographic characteristics as well as procedural and clinical events will be collected. The primary outcome measure is the occurrence of a major adverse cardiac event (MACE; defined as death, non-fatal myocardial-infarction, or need for repeat-revascularisation). Secondary outcome measures are the occurrence of major adverse cardiac and cerebrovascular events (MACCE; defined as death, non-fatal myocardial-infarction, repeat-revascularisation, or stroke), patient-reported quality of life, and health-economic data. Patient assessments will be performed during hospitalisation, at 1-month, 1-year, and annually thereafter to 5 years post-CABG. Events will be adjudicated by an independent clinical events committee. This European, multi-institutional registry will provide detailed insights into clinical outcome associated with DuraGraft. DISCUSSION: This European, multi-institutional registry will provide detailed insights into clinical outcome associated with the use of DuraGraft. Beyond that, and given the comprehensive data sets comprising of patient, procedural, and graft parameters that are being collected, the registry will enable for multiple subgroup analyses targeting focus groups or specific clinical questions. These may include analysis of subpopulations such as patients with diabetes or multimorbid high-risk patients (patient level), evaluation of relevance of harvesting technique including endoscopic versus open conduit harvesting (procedural level), or particular graft-specific aspects (conduit level). TRIAL REGISTRATION: ClinicalTrials.gov NCT02922088 . Registered October 3, 2016. ETHICS AND DISSEMINATION: The regional ethics committees have approved the registry. Results will be submitted for publication.


Assuntos
Ponte de Artéria Coronária/métodos , Doença da Artéria Coronariana/cirurgia , Artéria Torácica Interna/transplante , Soluções para Preservação de Órgãos/uso terapêutico , Artéria Radial/transplante , Veia Safena/transplante , Grau de Desobstrução Vascular , Idoso , Endoscopia , Endotélio Vascular , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Mortalidade , Infarto do Miocárdio/epidemiologia , Estudos Prospectivos , Qualidade de Vida , Sistema de Registros , Reoperação/estatística & dados numéricos , Acidente Vascular Cerebral/epidemiologia , Resultado do Tratamento
8.
Nat Biotechnol ; 37(10): 1131-1136, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31501557

RESUMO

The inability to preserve vascular organs beyond several hours contributes to the scarcity of organs for transplantation1,2. Standard hypothermic preservation at +4 °C (refs. 1,3) limits liver preservation to less than 12 h. Our group previously showed that supercooled ice-free storage at -6 °C can extend viable preservation of rat livers4,5 However, scaling supercooling preservation to human organs is intrinsically limited because of volume-dependent stochastic ice formation. Here, we describe an improved supercooling protocol that averts freezing of human livers by minimizing favorable sites of ice nucleation and homogeneous preconditioning with protective agents during machine perfusion. We show that human livers can be stored at -4 °C with supercooling followed by subnormothermic machine perfusion, effectively extending the ex vivo life of the organ by 27 h. We show that viability of livers before and after supercooling is unchanged, and that after supercooling livers can withstand the stress of simulated transplantation by ex vivo normothermic reperfusion with blood.


Assuntos
Temperatura Baixa , Fígado/fisiologia , Preservação de Órgãos/métodos , Humanos , Soluções para Preservação de Órgãos , Perfusão , Sobrevivência de Tecidos
10.
Ann R Coll Surg Engl ; 101(8): 609-616, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31508984

RESUMO

INTRODUCTION: Hypothermic machine perfusion, an organ preservation modality, involves flow of chilled preservation fluid through an allograft's vasculature. This study describes a simple, reproducible, human model that allows for interrogation of flow effects during ex vivo organ perfusion. MATERIALS AND METHODS: Gonadal veins from deceased human renal allografts were subjected to either static cold storage or hypothermic machine perfusion for up to 24 hours. Caspase-3, Krüppel-like factor 2 expression and electron microscopic analysis were compared between 'flow' and 'no-flow' conditions, with living donor gonadal vein sections serving as negative controls. RESULTS: The increase in caspase-3 expression was less pronounced for hypothermic machine-perfused veins compared with static cold storage (median-fold increase 1.2 vs 2.3; P < 0.05). Transmission electron microscopy provided ultrastructural corroboration of endothelial cell apoptosis in static cold storage conditions. For static cold storage preserved veins, Krüppel-like factor 2 expression diminished in a time-dependent manner between baseline and 12 hours (P < 0.05) but was abrogated and reversed by hypothermic machine perfusion (P < 0.05). CONCLUSIONS: Our methodology is a simple, reproducible and successful model of ex vivo perfusion in the context of human organ preservation. To demonstrate the model's utility, we establish that two widely used markers of endothelial health (caspase-3 and Krüppel-like factor 2) differ between the flow and no-flow conditions of the two predominant kidney preservation modalities. These findings suggest that ex vivo perfusion may mediate the induction of a biochemically favourable endothelial niche which may contribute tohypothermic machine perfusion's association with improved renal transplantation outcomes.


Assuntos
Transplante de Rim/métodos , Rim/irrigação sanguínea , Modelos Biológicos , Soluções para Preservação de Órgãos/farmacocinética , Preservação de Órgãos/métodos , Apoptose , Biomarcadores/metabolismo , Cadáver , Caspase 3/metabolismo , Temperatura Baixa , Endotélio Vascular/metabolismo , Humanos , Rim/metabolismo , Rim/ultraestrutura , Fatores de Transcrição Kruppel-Like/metabolismo , Microscopia Eletrônica , Perfusão/métodos , Veias/metabolismo , Veias/ultraestrutura
11.
Mol Med Rep ; 20(3): 2101-2110, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31257518

RESUMO

Ischemia­reperfusion injury (IRI) is a notable cause of tissue damage during surgical procedures and a major risk factor in graft dysfunction in liver transplantation. Livers obtained from donors after circulatory death (DCD) are prone to IRI and toll­like receptor 4 (TLR4) serves a prominent role in the inflammatory response associated with DCD liver IRI. The present study was designed to investigate whether TAK242, a specific TLR4 inhibitor, improves hepatic IRI following a DCD graft and to investigate its underlying protective mechanisms. Male Sprague­Dawley rats were randomized into 4 groups: Control, TAK242, DCD and DCD+TAK242 groups. Rats were pretreated with TAK242 or its vehicle for 30 min, then the livers were harvested without warm ischemia (control group and TAK242 group) or with warm ischemia in situ for 30 min. The livers were stored in cold University of Wisconsin solution for 24 h and subsequently perfused for 60 min with an isolated perfused rat liver system. Rat liver injury was evaluated thereafter. When compared with the DCD group, DCD livers with TAK242 pretreatment displayed significantly improved hepatic tissue injury and less tissue necrosis (P<0.05). Compared with DCD livers, mechanistic experiments revealed that TAK242 pretreatment alleviated mitochondrial dysfunction, reduced reactive oxygen species and malondialdehyde levels and inhibited apoptosis. Additionally, TAK242 significantly inhibited the IRI­associated inflammatory response, indicated by the decreased expression of TLR4, interleukin (IL)­1ß, IL­6 and cyclooxygenase 2 at the mRNA and protein levels (P<0.05). TAK242 ameliorates DCD liver IRI via suppressing the TLR4 signaling pathway in rats. The results of the present study have revealed that TAK242 pretreatment harbors a potential benefit for liver transplantation.


Assuntos
Anti-Inflamatórios/farmacologia , Fígado/efeitos dos fármacos , Preservação de Órgãos/métodos , Traumatismo por Reperfusão/imunologia , Sulfonamidas/farmacologia , Receptor 4 Toll-Like/antagonistas & inibidores , Adenosina/farmacologia , Alopurinol/farmacologia , Animais , Glutationa/farmacologia , Insulina/farmacologia , Fígado/imunologia , Fígado/patologia , Fígado/ultraestrutura , Transplante de Fígado , Masculino , Soluções para Preservação de Órgãos/farmacologia , Rafinose/farmacologia , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/imunologia , Isquemia Quente
12.
J Card Surg ; 34(10): 969-975, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31332833

RESUMO

OBJECTIVES: Cold crystalloid cardioplegia for donor heart harvesting and cold ischemic storage conditions during the transportation is the standard of care during heart transplantation procedure. Organ care system (OCS) was introduced for more prolonged and reliable ex vivo organ management. This study evaluated the two different techniques used for myocardial preservation during the procurement and transportation of the heart using the OCS. METHODS: We performed prospective analysis of 43 patients with heart failure undergoing heart transplantation and using the OCS for donor organ transport. Donor hearts were arrested using blood cardioplegia and conditioning (n = 30) or standard Custodiol (SC) solution ( n = 13). Perfusion and cardiac function parameters were continuously monitored while the donor hearts were perfused in the OCS. Impact of preservation techniques on biochemical parameters and clinical outcomes were evaluated. RESULTS: All donor hearts had stable perfusion and lactate characteristics in the OCS, with similar measures between the two groups at the beginning of the ex vivo perfusion. Ex vivo heart perfusion mean ending concentration of Interleukin (IL)-6 and IL-8 was significantly lower in the blood cardioplegia group compared to the standard care group. Clinical outcomes were comparable between the two groups of patients. CONCLUSIONS: The use of blood cardioplegia and conditioning could be a safe method for myocardial protection in distant procurement and preservation of donor hearts in the OCS.


Assuntos
Parada Cardíaca Induzida/métodos , Insuficiência Cardíaca/cirurgia , Transplante de Coração/métodos , Preservação de Órgãos/métodos , Perfusão/métodos , Doadores de Tecidos , Adulto , Feminino , Seguimentos , Glucose/farmacologia , Humanos , Masculino , Manitol/farmacologia , Soluções para Preservação de Órgãos/farmacologia , Cloreto de Potássio/farmacologia , Procaína/farmacologia , Estudos Prospectivos
13.
Cornea ; 38(10): 1314-1321, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31335527

RESUMO

PURPOSE: To evaluate a new corneal cold storage medium including an antimycotic tablet (Kerasave, AL.CHI.MI.A. S.r.l.). METHODS: Kerasave and tryptone soy broth (control) were inoculated with 10 and 10 colony-forming units (CFU)/mL of 6 Candida isolates (Candida albicans [n = 4], Candida tropicalis [n = 1], and Candida glabrata [n = 1]). Minimum inhibitory concentrations (MICs) were determined using amphotericin B Etest strips. Sterile porcine corneas contaminated with 10 CFU/mL of each isolate were incubated in Kerasave and control at 4°C. Growth rate and Log10 reduction at 4°C at different time intervals were determined for liquid samples and tissue homogenates. Kerasave biocompatibility was assessed according to ISO 10993-5 and ISO 10993-10. RESULTS: No C. albicans or C. tropicalis colonies were recovered from Kerasave inoculated with 10 CFU/mL after incubation for 3 days at 4°C. C. glabrata was inhibited but not killed after 3 days at 4°C. Four of the 6 strains contaminated with 10 CFU/mL demonstrated a significant ≥ 3 Log10 reduction in media and tissue homogenates within 5 days as compared to controls (p < 0.01). Amphotericin B MICs ranged from 0.19 to 0.38 µg/mL for C. albicans (n = 3) and C. tropicalis (n = 1). C. glabrata showed reduced susceptibility (0.5 µg/mL) and 1 C. albicans was resistant to amphotericin B (≥ 1 µg/mL). Kerasave was not cytotoxic, irritating, or sensitizing according to the ISO standards. CONCLUSIONS: Kerasave showed high antifungal efficacy against susceptible fungal strains at 4°C in the presence and absence of corneal tissue. Resistant strains to amphotericin B were not eliminated by Kerasave. Kerasave is not cytotoxic, irritating, or sensitizing.


Assuntos
Anfotericina B/farmacologia , Candida/efeitos dos fármacos , Candidíase/tratamento farmacológico , Córnea/efeitos dos fármacos , Infecções Oculares Fúngicas/tratamento farmacológico , Ceratite/tratamento farmacológico , Soluções para Preservação de Órgãos/farmacologia , Animais , Antifúngicos/farmacologia , Candida/isolamento & purificação , Candidíase/diagnóstico , Candidíase/microbiologia , Córnea/diagnóstico por imagem , Modelos Animais de Doenças , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/microbiologia , Ceratite/diagnóstico , Ceratite/microbiologia , Testes de Sensibilidade Microbiana , Preservação de Órgãos/métodos , Suínos
14.
Transpl Infect Dis ; 21(5): e13135, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31246353

RESUMO

BACKGROUND: Owing to organ shortage, transplantation of organs from HCV (hepatitis C virus) viremic donors into HCV negative individuals is getting more and more accepted. However, transmission of HCV to the host is nearly universal. Until now it is unknown if preservation solutions (PS) might alter infectivity and stability of HCV in the transplant setting. Therefore, seven different preservation solutions (PS) with variable composition were tested in vitro for their direct anti- and proviral effects on HCV. METHODS: In vitro grown HCV based on the JFH-1 isolate was used to characterize the effect of seven different PS on the HCV replication cycle including HCV attachment, entry, replication, and assembly. In addition, HCV stability in PS was tested. RESULTS: Overall, 6/7 PS enhanced HCV infectivity: IGL-1 increased HCV attachment and entry, UW Belzer and Perfadex boosted HCV entry. Production of novel viral particles was enhanced in HTK, UW Belzer, and IGL-1. In contrast, viral replication was significantly reduced in HTK solution while all other PS had no effect on HCV RNA replication. HCV was significantly more stable in HTK solution. Euro Collins was the only PS that did not support HCV infectivity in cell culture. None of the used PS showed cytotoxic effects. CONCLUSION: Our data indicate that HCV infectivity and stability is maintained by several PS.


Assuntos
Hepacivirus/efeitos dos fármacos , Soluções para Preservação de Órgãos/farmacologia , Replicação Viral/efeitos dos fármacos , Linhagem Celular , Hepacivirus/fisiologia , Humanos , Ligação Viral/efeitos dos fármacos , Internalização do Vírus/efeitos dos fármacos
15.
J Indian Soc Pedod Prev Dent ; 37(2): 140-145, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31249176

RESUMO

Aims: The aim of the study is to evaluate the efficacy of neem and turmeric as storage media in maintaining periodontal ligament (PDL) cell viability. Materials and Methods: Ninety premolar extracted teeth were randomly selected and divided into three groups, namely milk as a control group and neem and turmeric as experimental groups. The teeth were placed in one of the three storage media for 30 min. Scrapped PDL fibers were collected in Falcon tubes, incubated, and centrifuged for 5 min at 800 rpm. Obtained PDL cells were stained with trypan blue, observed, and counted in a hemocytometer under microscope, which was followed by the calculation of percentages of viable cells. One-way ANOVA was applied for comparison between different groups, and Tukey's test was applied for pair-wise comparison. Results: Mean percentage of viable cells in milk was 89.98 ± 4.11%, whereas in neem and turmeric extracts, it was 88.00 ± 5.85% and 81.63 ± 7.12%, respectively. There was a significant difference between all the three storage media for the viable PDL cells (P = 0.001). Intergroup comparison of the different storage media showed that there was a statistically highly significant difference between milk and turmeric (P ≤ 0.001). There was a statistically significant difference in between turmeric and neem (P ≤ 0.531) for the viable cell percentage, with neem being better storage medium than the turmeric. Conclusion: Within the limitations of the current study, it can be concluded that neem is as efficient as milk in maintaining the PDL cell viability. Turmeric, though is an efficient storage medium, was not as efficient as milk and neem.


Assuntos
Azadirachta , Soluções para Preservação de Órgãos , Avulsão Dentária , Animais , Sobrevivência Celular , Curcuma , Leite , Ligamento Periodontal , Extratos Vegetais
16.
Int J Mol Sci ; 20(13)2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31252560

RESUMO

Advanced preservation injury (PI) after intestinal transplantation has deleterious short- and long-term effects and constitutes a major research topic. Logistics and costs favor rodent studies, whereas clinical translation mandates studies in larger animals or using human material. Despite diverging reports, no direct comparison between the development of intestinal PI in rats, pigs, and humans is available. We compared the development of PI in rat, porcine, and human intestines. Intestinal procurement and cold storage (CS) using histidine-tryptophan-ketoglutarate solution was performed in rats, pigs, and humans. Tissue samples were obtained after 8, 14, and 24 h of CS), and PI was assessed morphologically and at the molecular level (cleaved caspase-3, zonula occludens, claudin-3 and 4, tricellulin, occludin, cytokeratin-8) using immunohistochemistry and Western blot. Intestinal PI developed slower in pigs compared to rats and humans. Tissue injury and apoptosis were significantly higher in rats. Tight junction proteins showed quantitative and qualitative changes differing between species. Significant interspecies differences exist between rats, pigs, and humans regarding intestinal PI progression at tissue and molecular levels. These differences should be taken into account both with regards to study design and the interpretation of findings when relating them to the clinical setting.


Assuntos
Mucosa Intestinal/transplante , Preservação de Órgãos/efeitos adversos , Transplantes/normas , Adolescente , Adulto , Animais , Caspase 3/genética , Caspase 3/metabolismo , Conexinas/genética , Conexinas/metabolismo , Criopreservação/métodos , Feminino , Humanos , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Masculino , Pessoa de Meia-Idade , Preservação de Órgãos/métodos , Soluções para Preservação de Órgãos/efeitos adversos , Soluções para Preservação de Órgãos/química , Ratos , Ratos Sprague-Dawley , Especificidade da Espécie , Suínos
17.
Folia Med Cracov ; 59(1): 101-114, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31180079

RESUMO

OBJECTIVES: To evaluate the properties of natural sweetener solutions in whole organ preservation and assess their influence on the dimension, weight and shape of cardiac tissue samples in stated time intervals, up to a one-year period of observation. BACKGROUND: Tissue fixation is essential for biological sample examination. Many negative toxic effects of formaldehyde-based fixatives have forced us to seek alternatives for formaldehyde based solutions. It has been demonstrated that natural sweeteners can preserve small tissue samples well and that these solutions can be used in histopathological processes. However, their ability to preserve whole human organs are unknown. METHODS: A total of 30 swine hearts were investigated. Three study groups (n = 10 in each case) were formed and classified on the type of fixative: (1) 10% formaldehyde phosphate-buffered solution (FPBS), (2) 10% alcohol-based honey solution (ABHS), (3) 10% water-based honey solution (WBHS). Samples were measured before fixation and in the following time points: 24 hours, 72 hours, 168 hours, 3 months, 6 months and 12 months. RESULTS: The WBHS failed to preserve heart samples and decomposition of tissues was observed one week after fixation. In half of the studied parameters, the ABHS had similar modifying tendencies as compared to FPBS. e overall condition of preserved tissue, weight, left ventricular wall thickness, right ventricular wall thickness and the diameter of the papillary muscle differed considerably. CONCLUSIONS: The ABHS may be used as an alternative fixative for macroscopic studies of cardiac tissue, whereas the WBHS is not suited for tissue preservation.


Assuntos
Etanol , Fixadores , Formaldeído , Coração/anatomia & histologia , Mel , Soluções para Preservação de Órgãos , Animais , Tamanho do Órgão , Suínos
18.
Anim Reprod Sci ; 207: 95-106, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31208848

RESUMO

This study was conducted to assess the effects of commercial extenders and storage temperature on dromedary camel sperm quality during liquid preservation. In Experiment 1, ejaculates (n = five males; replicated seven times) were split and diluted with synthetic (OPTIXcell, EquiPlus, INRA96, Bioxcell or AndroMed; Experiment 1a) or egg-yolk based (Biladyl, Green buffer or Triladyl; Experiment 1b) extenders and stored for 48 h at 4 °C. In Experiment 2, split ejaculates (n = five males; replicated six times) were used to directly compare Green buffer, OPTIXcell and Triladyl extenders over 48 h of storage at 4 °C. Ejaculates collected in Experiment 3 (n = five males; replicated five times) were diluted with Green buffer or Triladyl before chilled storage for 48 h at 4 or 15 °C. Sperm kinematics, viability and acrosome integrity were assessed during liquid storage. In Experiment 1a, there was the greatest total sperm motility (TM) in the OPTIXcell group following 24 and 48 h of storage, while in Experiment 1b, there was the greatest TM after 48 h of storage with Triladyl and Green buffer. In Experiment 2, there were greater TM and viable acrosome intact spermatozoa in the Triladyl and Green buffer than with OPTIXcell group. In Experiment 3, there was a greater TM in the Triladyl than Green buffer group at 24 and 48 h of storage regardless of storage temperature (which had no effect on sperm quality). In conclusion, camel sperm have greater viability when preserved in liquid form for 48 h following dilution with Triladyl and storage at either 4 or 15 °C.


Assuntos
Camelus , Soluções para Preservação de Órgãos/farmacologia , Refrigeração , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Sêmen/efeitos dos fármacos , Animais , Tampões (Química) , Sobrevivência Celular/efeitos dos fármacos , Gema de Ovo/fisiologia , Soluções Isotônicas/farmacologia , Masculino , Soluções para Preservação de Órgãos/química , Refrigeração/métodos , Refrigeração/veterinária , Análise do Sêmen , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Temperatura Ambiente , Fatores de Tempo
19.
Transpl Infect Dis ; 21(4): e13123, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31165536

RESUMO

PURPOSE: This study investigated the donor characteristics of methicillin-resistant Staphylococcus aureus (MRSA) contamination in storage medium before transfer of corneas to preservation medium for corneal transplantation, in order to identify donor characteristic risk factors for MRSA contamination. METHODS: This retrospective, cross-sectional study was performed using Juntendo Eye Bank records for all corneal transplantation procedures. Storage medium (EP-II® ) cultures for right eyes were included for the period between July 2008 and December 2017. The following donor characteristics were collected: age, sex, cause of death, history of cataract surgery, death-to-enucleation interval, death-to-preservation interval, and endothelial cell density (ECD). Donor characteristics were compared between MRSA and non-MRSA contamination groups. Odds ratios (ORs) for donor-related risk factors for MRSA contamination were determined using logistic regression. RESULTS: In total, 370 storage medium samples were examined; 222 were positive for bacterial cultures (60.0%) and 36 were MRSA-positive (9.7%). Donor age was significantly higher in the MRSA contamination group than in the non-MRSA contamination group (86.1 ± 9.5 years vs 75.9 ± 15.9 years, P < 0.001). Univariate logistic regression analysis showed that MRSA contamination risk factors were older age (OR = 1.07; 95% confidence interval [95% CI]: 1.03-1.11) and decreased ECD (OR = 0.9993; 95% CI: 0.9986-0.9992). The fully adjusted OR for every year of age as a risk factor for MRSA contamination was 1.07 (95% CI: 1.03-1.11). CONCLUSIONS: Aging was a risk factor for MRSA contamination in storage medium. Careful pre-banking assessment of elderly donor corneas is needed to prevent intractable postoperative MRSA infection.


Assuntos
Transplante de Córnea , Bancos de Olhos/normas , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Soluções para Preservação de Órgãos , Infecções Estafilocócicas/prevenção & controle , Idoso , Idoso de 80 Anos ou mais , Córnea , Estudos Transversais , Infecções Oculares Bacterianas/prevenção & controle , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Estudos Retrospectivos , Fatores de Risco , Manejo de Espécimes , Doadores de Tecidos , Obtenção de Tecidos e Órgãos
20.
J Reprod Dev ; 65(4): 353-359, 2019 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-31118350

RESUMO

Freeze-drying of spermatozoa is a convenient and safe method to preserve mammalian genetic material without the use of liquid nitrogen or a deep freezer. However, freeze-dried spermatozoa (FD sperm) are not frequently used because of the low success rate of offspring after intracytoplasmic spermatozoa injection (ICSI). In this study, we determined the optimal concentration and a point of action of trehalose as a protectant for the preservation of FD sperm from different mouse strains at room temperature (RT). Although trehalose demonstrated no potential to protect the FD sperm of ICR mice against the freeze-drying procedure itself, the blastocyst rate was significantly improved when FD sperm was preserved for more than 1 month at RT (56-63% vs. 29% without trehalose). The optimal concentration of trehalose was 0.5 M. Importantly, remarkable results were obtained when spermatozoa of inbred mouse strains (C57BL/6N, C3H/He, and 129/Sv) were used, and many offspring were obtained when FD sperm that was preserved for 3 months at RT (26-28% vs. 6-11% of without trehalose) was used. However, when DNA damage in FD sperm was examined by gamma-H2Ax assays, it was found that trehalose failed to protect the FD sperm from DNA damage. These results suggest that trehalose has the potential to protect other sperm factors rather than sperm DNA during preservation at RT for longer periods and trehalose is more effective for inbred mouse strains.


Assuntos
Preservação do Sêmen/métodos , Espermatozoides , Trealose/farmacologia , Animais , Feminino , Liofilização/métodos , Liofilização/veterinária , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Endogâmicos ICR , Soluções para Preservação de Órgãos/farmacologia , Gravidez , Taxa de Gravidez , Preservação do Sêmen/veterinária , Injeções de Esperma Intracitoplásmicas
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