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1.
BMC Ophthalmol ; 22(1): 137, 2022 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-35337290

RESUMO

BACKGROUND: To evaluate the surface quality and thickness uniformity of lamellar donor grafts using an optional surgical technique called reversed manual dissection (RMD) in porcine corneas. METHODS: Twenty-four paired porcine corneas (48 eyes) were numbered 1 to 24 and divided into 6 groups. All left corneas were assigned to conventional manual dissection (CMD), and all right corneas were assigned to RMD. Each group contained 8 corneas. For Groups I, II, and III, 30, 50, and 70% of the entire corneal thickness was dissected using CMD. For groups IV, V, and VI, 70, 50, and 30% of the entire corneal thickness was dissected using RMD. The residual stromal thickness was examined by anterior segment optical coherence tomography (ASOCT) to assess the thickness uniformity and scanning electron microscopy (SEM) to assess the surface quality. RESULTS: The thickness uniformity of the lamellar grafts between each paired group was not significantly different (p > 0.05). The qualitative surface roughness grading (QiSR) evaluated by masked observers through SEM was significantly higher in the RMD groups (p < 0.001). The quantitative surface roughness grading (QnSR) acquired from the Mountains software was significantly lower in the RMD groups (p < 0.001). CONCLUSIONS: RMD is an optional surgical technique for obtaining porcine lamellar grafts. The thickness uniformity of RMD is comparable to that of CMD, and a smoother surface with fewer ridges and roughness is achieved compared to CMD.


Assuntos
Córnea , Substância Própria , Animais , Córnea/cirurgia , Substância Própria/cirurgia , Microscopia Eletrônica de Varredura , Projetos Piloto , Suínos , Tomografia de Coerência Óptica
2.
Graefes Arch Clin Exp Ophthalmol ; 257(1): 135-141, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30506095

RESUMO

PURPOSE: Donor-to-host transmission of infectious agents is a rare but well-recognised complication of corneal transplantation and may carry a grave visual prognosis. In this case series, we describe the clinical features and risk factors of using culture-positive donor corneas for transplantation. METHODS: Retrospective chart review of a series of patients who underwent either penetrating keratoplasty (PK) or Descemet's stripping automated endothelial keratoplasty (DSAEK) with positive microbiology cultivation during routine assessment of donor corneal tissue obtained at the time of surgery. Donor and recipient characteristics, tissue preparation and surgical parameters, clinical signs and outcomes were registered. RESULTS: Eleven patients who received culture-positive corneal grafts were identified: six with Candida, three with Gram-positive bacteria and two with Gram-negative bacteria. Three patients developed clinical keratitis after routine DSAEK using corneas contaminated with Candida species. The median death-to-preservation time (DPT) of these three donor corneas was 18.08 (range 18.08 to 20.90) h, while in the remaining eight donors, it was 12.27 (range 9.32 to 20.47) h. Despite the initiation of antifungal treatment, all three cases required explantation of the graft and a subsequent re-DSAEK. CONCLUSIONS: The use of donor corneas that are culture-positive for Candida carries a risk for developing postoperative keratitis and the risk may be higher in DSAEK. Unlike the cold storage technique employed for donor corneas described in this case series, organ culture technique requires microbiological screening and supplementation of an antifungal agent which may reduce the risk of donor-to-host transmission of fungal infection.


Assuntos
Córnea/microbiologia , Infecções Oculares Bacterianas/transmissão , Infecções Oculares Fúngicas/transmissão , Ceratite/microbiologia , Ceratoplastia Penetrante , Infecção da Ferida Cirúrgica/microbiologia , Doadores de Tecidos , Idoso , Idoso de 80 Anos ou mais , Córnea/cirurgia , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Fúngicas/microbiologia , Feminino , Sobrevivência de Enxerto , Humanos , Ceratite/etiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Infecção da Ferida Cirúrgica/etiologia
3.
Histochem Cell Biol ; 150(5): 567-574, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30094468

RESUMO

Organ explant cultures are well-established in vitro models that are used to study normal cell biological and regeneration processes as well as carcinogenesis. Primary urothelial cultures from bladder mucosa explants are highly differentiated and are thus broadly used as in vitro experimental equivalents of native urothelial tissue. Since experiments on differentiated urothelial cultures from bladder mucosa explants currently allow only a single use of explants, establishment of sufficient quantities of cultures requires large numbers of sacrificed animals. There is thus a great need for a cheaper approach with less ethical dilemmas. Herein, we demonstrate that mouse bladder mucosa explants can be reused. Reused explants produce outgrowths with highly differentiated urothelia, just like primary explants. Even after being recycled ten times, urothelial outgrowths have the supramolecular and ultrastructural features that are comparable to the native urothelium. Ten times reused explants produce superficial urothelial cells that express uroplakins in the apical plasma membrane, claudin-8 in the tight junctions, and have a subapical network of cytokeratin 20. Basal urothelial cells in urothelial outgrowths of ten times reused explants express p63 which indicates that these urothelial outgrowths have a persistent proliferative capacity. Using our approach, one can perform experiments that were previously not feasible due to low quantities of donor tissue. The method also offers opportunity for effective use of scarce healthy human urothelial tissue.


Assuntos
Diferenciação Celular , Mucosa/citologia , Bexiga Urinária/citologia , Urotélio/citologia , Animais , Técnicas de Cultura de Células/métodos , Células Cultivadas , Reutilização de Equipamento , Imunofluorescência , Masculino , Camundongos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão
5.
Surg Radiol Anat ; 40(2): 199-206, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29204678

RESUMO

PURPOSE: The greater and lesser palatine nerves and vessels supply the hard and soft palates, and the roots of these vessels and nerves run through a bony structure. However, the arrangement of blood vessels in the maxilla requires attention during clinical treatments, but detailed morphological information about changes in the greater and lesser palatine arteries and nerves during aging is unavailable. We therefore need detailed investigations of the morphology of the donor cadaver palatine using cone-beam computed tomography (CBCT) and macroscopic observations. METHODS: We investigated 72 donor cadavers using macroscopic segmentation and CBCT. The results' analysis examined differences in skull measurement parameters and differences between dentate and edentulous cases. RESULTS: The greater palatine artery and nerve showed different macroscopic arrangements in dentate and edentulous cadavers. We also classified three types of bony structures of the nerve and vessel roots in the molar regions of the palatine using CBCT images: the shallow groove, deep groove, and flat groove. The deep groove is the deepest of the three and is remarkable in edentulous elderly cadavers. CONCLUSION: This study of macroscopic and CBCT data provides information useful for planning dental implant surgeries and autogenous bone harvesting.


Assuntos
Tomografia Computadorizada de Feixe Cônico , Palato Duro/irrigação sanguínea , Palato Duro/inervação , Palato Mole/irrigação sanguínea , Palato Mole/inervação , Adulto , Idoso , Cadáver , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Palato Duro/diagnóstico por imagem , Palato Mole/diagnóstico por imagem
6.
Int Ophthalmol ; 38(4): 1669-1679, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28688026

RESUMO

PURPOSE: To study the indications and variables correlating with graft survival in optical penetrating keratoplasty in a tertiary care hospital in north India. PARTICIPANTS: All patients who underwent optical keratoplasty, except those undergoing lamellar grafts, tectonic grafts transplants and penetrating keratoplasty done for therapeutic purposes were included in the study. Patients with follow-up less than 2 years were excluded from the study. MATERIALS AND METHODS: Data were obtained by reviewing the records of 101 patients who underwent optical penetrating keratoplasty from 2008 to 2013 for various indications. RESULTS: Out of 101 patients who underwent optical penetrating keratoplasty, 71 were males and 30 were females. The mean age of the recipient was 48.53 years with range 1-82 years. The main indications were previous failed graft (29.7%), healed keratitis except HSV (15.8%), pseudophakic or aphakic bullous keratopathy (14.8%), corneal dystrophies/degenerations (12.9%), adherent leucoma (9.9%), post-HSV scars (8.9%), and others like anterior staphyloma, congenital corneal opacities, buphthalmos and keratoconus (7.9%). The graft survival rate was 67.33% at 1-year follow-up and 59.4% at 2-year follow-up. The mean survival time of the grafts was 22.42 months. The mean time for graft failure was 7.12 (±0.9) months. CONCLUSION: One- and 2-year survival of grafts at our center is lower as compared to western studies, probably due to higher percentage of poor prognosis indications for surgery and a relative scarcity of excellent-quality donor corneas.


Assuntos
Doenças da Córnea/cirurgia , Sobrevivência de Enxerto , Ceratoplastia Penetrante , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Células Endoteliais/citologia , Feminino , Humanos , Índia , Lactente , Estimativa de Kaplan-Meier , Ceratoplastia Penetrante/métodos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Técnicas de Sutura , Centros de Atenção Terciária , Acuidade Visual , Adulto Jovem
7.
BMC Ophthalmol ; 17(1): 204, 2017 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-29149876

RESUMO

BACKGROUND: To evaluate main features of donor tissue that may influence clinical outcome or complication rate after deep anterior lamellar keratoplasty (DALK). METHODS: Donor tissue parameters of 84 consecutive corneal donor grafts used for big-bubble DALK surgery between June 2011 and December 2014 in 84 eyes of 84 patients with disorders of anterior corneal stroma were correlated to clinical outcome parameters of recipient eyes 12 months after surgery and 3 months after total suture removal. Main donor tissue parameters included age), post-mortem time, overall preservation time, preservation time after split and prior to transplantation, and preservation technique. Clinical outcome parameters included best spectacle corrected visual acuity (BSCVA), central corneal thickness (CCT), endothelial cell density (ECD) and complication rates. Pearson's correlation, linear regression analysis for clinical outcome parameter and logistic regression analysis for postsurgical complication rates were applied. RESULTS: Corneal donors were mean aged 67.4 ± 12.5 years with a post-mortem time of 20.7 ± 14.7 h and ECD of 2641.0 ± 362.8 cells/mm2. Overall preservation time was 16.3 ± 6.3 days. Recipients showed mean BSCVA 12 months postoperatively of 0.60 ± 0.36 logMAR, endothelial cell loss was 4 ± 16%, and central corneal thickness was 571.7 ± 54.2 µm. 3 months after total removal of sutures, BSCVA was 0.20 ± 0.10 logMAR, endothelial cell loss was 17 ± 24%, and central corneal thickness was 590.9 ± 55.5 µm. Loosening of sutures occurred in 20%, and Descemet detachment in 16%. None of the clinical outcome parameters or complication rate after DALK showed a significant association with donor tissue parameters. CONCLUSIONS: Donor corneas, independent of excision techniques or preservation method, with donor age ≤ 88 years, post-mortem time ≤ 63 h, overall preservation time ≤ 14 days for cold storaged donor tissue and ≤35 days for organ culture, and preservation time after split prior to grafting ≤96 h, seem to be applicable as safe donor tissue for DALK surgery.


Assuntos
Doenças da Córnea/cirurgia , Ceratoplastia Penetrante/métodos , Doadores de Tecidos , Adulto , Fatores Etários , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Tempo para o Tratamento , Preservação de Tecido/métodos , Acuidade Visual
8.
Cell Tissue Bank ; 18(1): 119-128, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27878655

RESUMO

Scleral tissue has been in use in ophthalmology for many years although indications for use have varied. We retrospectively reviewed scleral transplant tissue requests over a 12 month period at a local eye bank and confirmed a small but significant demand for the use of scleral tissue. Iatrogenic surgical complications are the primary indication for use. Our understanding of the indications and outcomes of scleral graft procedures is derived from case reports and small cohort series. We reviewed the current literature on existing indications for its use and discuss the relative outcomes. To our knowledge this represents the first review of scleral transplant indications and further summarises usage rates in the Lions NSW Eye Bank which may provide practical information for those surgeons who use scleral tissue and Eye Banks who supply it.


Assuntos
Procedimentos Cirúrgicos Oftalmológicos/métodos , Esclera/transplante , Bancos de Olhos , Humanos , Oftalmologia/métodos , Preservação de Órgãos/métodos , Esclera/anatomia & histologia , Coleta de Tecidos e Órgãos/métodos
9.
Surg Radiol Anat ; 39(2): 177-184, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27177906

RESUMO

OBJECTIVE: Investigation of the computerized dimensional anatomic location of the greater palatine foramen (GPF) and lesser palatine foramens (LPF) is important indicating site to collect palatal donor tissue, reconstructioning the orofacial area of the oncology patient and applying the greater palatine nerve block anesthesia. The aim of this study is to determine a patient-friendly landmark and to specify the precise location of the GPF in order to standardise certain anatomical marks of safe neurovascular bundle. MATERIALS AND METHODS: 120 bony palates were examined to detect the position of the GPF and the LPF related to adjacent anatomical landmarks using a computer software program. The GPF was assessed regarding the position, the diameter and the distances between each foramen and the midline maxillary suture (MMS), the inner border of alveolar ridge (AR), posterior palatal border (PBB), and incisive foramen (IF). RESULTS: The GPF was identified as single in 81 %, double in 16 %, triple in 2 % and absent in 2 % of the specimens. The mean distances between the GPF and the MSS, the GPF and the AR, the GPF and the PPB, the GPF and the IF were 16, 4, 4, and 40 mm, respectively. In majority of the cases, the GPF was seen between the distal surfaces of the third maxillary molar (78 %). Single LPF was observed in 53.45 % of the skulls, two LPF were observed in 31 % of the skulls bilaterally and five LPF were rare in 2.1 % of the specimens. The LPF was most commonly at the junction of the palatine bone and the inner lamella of the pterygoid plate (71.9 %). CONCLUSIONS: This study made possible to investigate the variability of the GPF and the feasibility of the greater palatine neurovascular bundle, and to calculate the lengths of some parameters with the help of certain software. To collect the donor tissue of the neurovascular greater palatine network, each distance among the AR-GPF-PPB were equal to 4 mm. To estimate the possible length of the graft, the incision was made along the third and the second molar to the IF as 4 cm. The data we obtained within this study have been presented to help the surgeons avoid unexpected hemorrhage during the palatinal procedures such as posttraumatic dental reconstruction, maxillofacial tumor resections, palatal micro-implants, and dentofacial orthopedic surgery.


Assuntos
Maxila/anatomia & histologia , Nervo Maxilar/anatomia & histologia , Palato Duro/anatomia & histologia , Palato Duro/cirurgia , Adulto , Processo Alveolar/anatomia & histologia , Variação Anatômica , Simulação por Computador , Humanos , Dente Molar/anatomia & histologia , Bloqueio Nervoso/métodos , Procedimentos Cirúrgicos Ortognáticos/métodos , Palato Duro/irrigação sanguínea , Palato Duro/inervação , Procedimentos de Cirurgia Plástica/métodos
10.
BJU Int ; 113(1): 160-6, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24053725

RESUMO

OBJECTIVES: To evaluate deceased non-heart beating (DNHB) donors and deceased heart beating (DHB) brain-stem dead donors, as sources of viable urological tissue for use in biomedical research. To identify sources of viable human bladder tissue as an essential resource for cell biological research aimed at understanding human diseases of the bladder and for developing new tissue engineering and regenerative medicine strategies for bladder reconstruction. Typically, normal human urinary tract tissue is obtained from adult or paediatric surgical patients with benign urological conditions, but few surgical procedures yield useful quantities of healthy bladder tissue for research. PATIENTS AND METHODS: Research ethics committee approval was obtained for collection of donor bladder tissue. Consent for DHB donors was undertaken by the Donor Transplant Coordinators. Tissue Donor Coordinators were responsible for consent for DNHB donors and the retrieval of bladders was coordinated through the National Blood Service Tissue Banking Service. All retrievals were performed by practicing urologists and care was taken to maintain sterility and to minimise bacterial contamination. Two bladders were retrieved from DNHB donors and four were retrieved from DHB donors. RESULTS: By histology, DNHB donor bladder tissue exhibited marked urothelial tissue damage and necrosis, with major loss or absence of urothelium. No cell cultures could be established from these specimens, as the urothelial cells were not viable in primary culture. Bladder urothelium from DHB donors was intact, but showed some damage, including loss of superficial cells and variable separation from the basement membrane. All four DHB bladder specimens yielded viable urothelial cells that attached in primary culture, but cell growth was slow to establish and cultures showed a limited capacity to form a functional barrier epithelium and a propensity to senesce early. CONCLUSIONS: We have shown that normal human bladder urothelial cell cultures can be established and serially propagated from DHB donor bladders. However, our study suggests that rapid post-mortem changes to the bladder affect the quality and viability of the urothelium, rendering tissue from DNHB donors an inadequate source for urothelial cell culture. Our experience is that whereas patients are willing to donate surgical tissue for research, there is a barrier to obtaining consent from next of kin for retrieved tissues to be used for research purposes.


Assuntos
Pesquisa Biomédica , Técnicas de Cultura de Células/métodos , Doadores de Tecidos , Bexiga Urinária/citologia , Urotélio/citologia , Pesquisa Biomédica/tendências , Técnicas de Cultura de Células/tendências , Proliferação de Células , Células Cultivadas , Feminino , Humanos , Imuno-Histoquímica , Masculino
11.
Front Med (Lausanne) ; 10: 1210293, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37608828

RESUMO

Introduction: The purpose of this review is to consolidate and examine the available literature on the coronavirus disease 2019 pandemic and its effect on corneal transplantation and eye banking. Methods: A primary literature search was conducted using the PubMed (Medline) database with keywords and MeSH terms such as "corneal transplantation," "eye banks," "keratoplasty" and then were combined with COVID-19. Relevant articles through September 2022 were assessed and 25 articles were included in this review. Results: Donor tissue volumes declined globally during lockdown periods due to a lower number of referrals and tighter tissue screening guidelines. Rates of elective surgeries decreased in the lockdown period compared to respective periods in previous years. However, changes in rates of emergency procedures were not uniform across different regions. Moreover, rates of different elective corneal grafts [i.e., penetrating keratoplasty (PK), endothelial keratoplasty (EK), or anterior lamellar keratoplasty (ALK)] were affected differently with the pattern of change being dependent on region-specific factors. Conclusion: Both donor tissue volumes and rates of corneal transplant procedures were affected by lockdown restrictions. The underlying etiology of these changes differed by region. Examining the range of impact across many countries as well as the contributing factors involved will provide guidance for future global pandemics.

12.
F S Sci ; 2(3): 248-258, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35146457

RESUMO

OBJECTIVE: Ovarian tissue cryopreservation is one of the crucial options for fertility preservation. Transplantation of cryopreserved ovarian tissue was proven to restore ovarian endocrine function in patients with premature ovarian insufficiency. Ovaries from deceased donors potentially serve as an excellent and readily available tissue for the translational and basic research. In this study, we used ovaries obtained from 5 deceased donors aged 18-26 years, to evaluate the number and quality of ovarian follicles isolated before and after cryopreservation. DESIGN: Preclinical. SETTING: Academic biomedical research laboratory. PATIENTS: De-identified deceased human donors. INTERVENTIONS: Slow-freeze cryopreservation and thawing. MAIN OUTCOME MEASURES: Follicle count, follicle density, follicle viability using immunohistochemical staining (TUNEL). RESULTS: The follicle density negatively correlated with age in both cryopreserved/thawed and fresh group. A total of 2803 follicles from fresh and 1608 follicles from cryopreserved tissues were classified and analyzed using Hematoxylin and eosin staining. There was no significant difference in the percent of morphologically normal follicles between two groups. TUNEL assay indicated no higher DNA damage in the follicles and the stroma cells after cryopreservation. Morphologically normal preantral follicles were enzymatically isolated from both fresh and cryopreserved tissue with 88.51 ± 5.93% (mean ± SD) of the isolated follicles confirmed viable using LIVE/DEAD evaluation. CONCLUSIONS: Our results indicate the ovarian tissue from deceased donors maintain high quality after long time extracorporeal circulation and transportation from the hospital to the laboratory. High survival rate of follicles at different developmental stages suggested tolerance to the cryopreservation process. Human ovarian tissues obtained from deceased donors is an ample source tissue and can be applied to promoting research and future clinical applications.


Assuntos
Preservação da Fertilidade , Ovário , Criopreservação/métodos , Feminino , Preservação da Fertilidade/métodos , Congelamento , Humanos , Folículo Ovariano
13.
Indian J Ophthalmol ; 68(6): 1044-1053, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32461427

RESUMO

Purpose: To analyze the overall clinical outcomes of Descemet membrane endothelial keratoplasty (DMEK) in 600 consecutive cases. Methods: Retrospective, consecutive interventional case series operated by a single surgeon. Six hundred consecutive eyes of 524 patients with endothelial dysfunctions of different etiologies scheduled for DMEK were included in this study. All donor tissues were prepared by the operating surgeon during the procedure, using McCarey Kaufman medium or Cornisol-preserved cornea with endothelial cell density (ECD) of ≥2500 cells/mm2. Indications, postoperative best spectacle-corrected visual acuity (BSCVA), ECD, endothelial cell loss (ECL), and complications were analyzed postoperatively between 3 months and 2 years. Results: The commonest indication was post-cataract corneal edema/bullous keratopathy in 262 (43.7%) eyes followed by Fuchs' endothelial corneal dystrophy 218 (36.3%). Vision affected comorbidities were present in 91 (15.2%) eyes. In phakic eyes with cataract (222; 37%), DMEK was combined with cataract surgery (Triple-DMEK). BSCVA of ≥20/25 was achieved in 41.0%, 46.4%, 49.2%, and 48.7% of eyes at 3, 6, 12, and 24 months, respectively and stabilized at 6 months (P = 0.54). Mean ECD decreased from 2884 ± 178 cells/mm2 (n = 600) before surgery to 2223 ± 321 (n = 597), 2099 ± 354 (n = 524), 1918 ± 373 (n = 374), and 1772 ± 439 cells/mm2 (n = 158) at 3, 6, 12, and 24 months respectively. The corresponding mean ECL was 22.9 ± 11.4%, 27.2 ± 12.4%, 33.5 ± 13.0%, and 38.6 ± 14.3%, respectively (P < 0.05 for all-time points). The commonest complication was DM detachment in 59 (9.8%) eyes of which 23 (3.8%) eyes required rebubbling. Three (0.5%) eyes had primary graft failure. Endothelial rejection occurred in 7 (1.2%) eyes until the last follow-up. Conclusion: DMEK is a safe and effective procedure in different types of endothelial diseases with encouraging surgical and clinical outcomes. Complications are less and ECL percentage up to 2 years is acceptable.


Assuntos
Ceratoplastia Endotelial com Remoção da Lâmina Limitante Posterior , Distrofia Endotelial de Fuchs , Contagem de Células , Perda de Células Endoteliais da Córnea/diagnóstico , Lâmina Limitante Posterior/cirurgia , Endotélio Corneano , Distrofia Endotelial de Fuchs/cirurgia , Humanos , Estudos Retrospectivos , Acuidade Visual
14.
Anaesth Intensive Care ; 46(6): 601-607, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30447670

RESUMO

Current donor management practices target macrohaemodynamic parameters, but it is unclear if this leads to improvements in microvascular perfusion and tissue oxygenation; the latter may have more impact on organ status. In a recent preclinical study we determined that brain death impaired tissue perfusion and oxygen utilisation in swine while pharmacologic correction of these deficits improved organ function and reduced markers of tissue injury. As a first step in translating the preclinical findings, we conducted a prospective observational study to determine if there was an association between peripheral tissue oxygenation (measured by near-infrared spectroscopy) in deceased by neurological criteria human donors and the number of organs transplanted. In 60 donors, the mean time-weighted average of tissue oxygenation was 87.5% (standard deviation, SD, 5.2%) and the average number of organs transplanted was 3.5 (SD 2); there was a positive linear relationship between these two parameters. A 5% rise in tissue oxygenation was associated with an increase of 0.47 organs transplanted (95% confidence intervals 0.16 to 0.78) after adjusting for age (P=0.004). No such correlations were observed for the macrohaemodynamic or macro-oxygenation parameters (including arterial blood oxygenation). The results of this clinical trial are consistent with our preclinical work and support the postulate that targeting the microvasculature to improve tissue perfusion and tissue oxygen delivery in human donors has the potential to increase the quantity of organs suitable for transplant.


Assuntos
Sistema Digestório/metabolismo , Rim/metabolismo , Pulmão/metabolismo , Miocárdio/metabolismo , Oxigênio/metabolismo , Doadores de Tecidos/estatística & dados numéricos , Obtenção de Tecidos e Órgãos/estatística & dados numéricos , Adulto , Feminino , Humanos , Masculino , Microvasos/metabolismo , Pessoa de Meia-Idade , Estudos Prospectivos , Espectroscopia de Luz Próxima ao Infravermelho
15.
Transl Vis Sci Technol ; 7(6): 11, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30510855

RESUMO

PURPOSE: To optimize methods of preparing donor cornea tissue for Descemet's stripping automated endothelial keratoplasty (DSAEK), we compared five experimental conditions with different internal pressures and cutting systems. METHODS: The artificial anterior chamber internal pressure (IP) was set at 100 or 200 mm Hg. The microkeratome cut was performed with or without an artificial chamber pressurizer (ACP), using a CBm turbine (CBm) or one use-plus automated (OUP-A). Thirty human research corneas were divided into five groups, and compared after the cut with donor tissue quality parameters, including cutting depth, graft uniformity, cell evaluation, and smoothness of the stromal surface. RESULTS: The smallest variation in mean cut depth was observed in the condition, which had IP of 200 mm Hg used ACP and OUP-A. In experimental groups cut using CBm, significantly more consistent thicknesses were made at an IP of 200 than 100 mm Hg. There were no statistically significant differences among the groups in either endothelial cell density or cell viable assay results after cuts. Using an IP of 200 mm Hg with ACP and CBm produced the roughest stromal surface, and the roughness grading scores showed a positive correlation with the percentage of cut depth. CONCLUSIONS: An IP of 200 mm Hg was the best setting for DSAEK grafts with high predictability of cut depth and uniformity of graft thickness without endothelial cell damage. TRANSLATIONAL RELEVANCE: For successful DSAEK, it is recommended that a set internal pressure of 200 mm Hg be used during microkeratome cutting for donor tissue preparation.

16.
Acta Ophthalmol ; 95(4): e297-e306, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28133954

RESUMO

PURPOSE: To provide quantitative parameters for assessment of human donor corneal stroma by imaging stromal features of diseased and normal human corneas with full-field optical coherence microscopy (FFOCM), using confocal microscopy (CM) and histology as reference techniques. METHODS: Bowman's layer (BL) thickness and keratocyte density were assessed ex vivo in 23 human donor corneas and 27 human pathological corneas (keratoconus and other corneal disorders) with FFOCM, CM and histology. Stromal backscattering was assessed with FFOCM. Additionally, 10 normal human corneas were assessed in vivo with CM. RESULTS: In FFOCM, the logarithm of the normalized stromal reflectivity was a linear function of stromal depth (R2  = 0.95) in human donor corneas. Compared with keratoconus corneas, human donor corneas featured higher BL thickness (p = 0.0014) with lower coefficient of variation (BL-COV; p = 0.0002), and linear logarithmic stromal reflectivity with depth (higher R2 , p = 0.0001). Compared with other corneal disorders, human donor corneas featured lower BL-COV (p = 0.012) and higher R2 (p = 0.0001). Using the 95% confidence limits of the human donor cornea group, BL thickness < 6.5 µm (sensitivity, 57%; specificity, 100%), BL-COV > 18.6% (79%; 100%) and R2  < 0.94 (93%; 71%) were revealed as indictors of abnormal cornea. In CM, keratocyte density decreased with stromal depth (r = -0.56). The mean overall keratocyte density (cells/mm2 ) was 205 in human donor corneas, 244 in keratoconus, 176 in other corneal disorders and 386 in normal corneas. CONCLUSION: Full-field optical coherence microscopy (FFOCM) provides precise and reliable parameters for non-invasive assessment of human donor corneal stroma during storage, enabling detection of stromal disorders that could impair the results of keratoplasty.


Assuntos
Doenças da Córnea/cirurgia , Substância Própria/citologia , Transplante de Córnea , Microscopia Confocal/métodos , Doadores de Tecidos , Tomografia de Coerência Óptica/métodos , Adulto , Idoso , Substância Própria/transplante , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
17.
Methods Mol Biol ; 1501: 115-129, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27796949

RESUMO

The mammary gland is the only organ to undergo most of its development after birth and therefore particularly attractive for studying developmental processes. In the mouse, powerful tissue recombination techniques are available that can be elegantly combined with the use of different genetically engineered mouse models to study development and differentiation in vivo.In this chapter, we describe how epithelial intrinsic gene function can by discerned by grafting mammary epithelial cells of different genotypes to wild-type recipients. Either pieces of mammary epithelial tissue or dissociated mammary epithelial cells are isolated from donor mice and subsequently transplanted into recipients whose mammary fat pads were divested of their endogenous epithelium. This is followed by phenotypic characterization of the epithelial outgrowth either by fluorescence stereomicroscopy for the fluorescently marked grafts or carmine alum whole mount for the unmarked epithelia.


Assuntos
Células Epiteliais/fisiologia , Epitélio/fisiologia , Glândulas Mamárias Animais/fisiologia , Animais , Diferenciação Celular/fisiologia , Feminino , Camundongos , Fenótipo
18.
Acta Ophthalmol ; 91(6): 571-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22863151

RESUMO

PURPOSE: To evaluate donor demographics, trends in donor tissue procurement and tissue storage over a long period. METHODS: A retrospective, longitudinal, descriptive analysis was undertaken of data from the Hamburg Eye Bank Data Base (HEB-DB) that had been collected between 1981 and 2010. Data on 54 parameters of cornea donors [including clinical history, age, death cause, gender and death-to-explantation interval (DEI)] and of cultivated corneas (endothelial quality and development in culture, cultivation period, microbiological contamination) were retrieved. These data were analysed statistically, focusing on the historical development of the eye bank. RESULTS: At the time of retrieval (June 2010), the HEB-DB contained data on 10 943 corneas (5503 donors). Most donors were men (65%) and had died from cardiopulmonary (n = 801)/cerebral (n = 261) failure or as the result of a polytraumatic accident/suicide (n = 602). Within these years, donor age, DEI and storage time increased. The percentage of stored corneas suitable for transplantation displayed a variable but increasing trend; in 2007, almost 75% of the stored corneas were transplanted. Between 1995 and June 2010, the median microbiological contamination rate was 5.3%. A change in the procurement procedure from enucleation to corneoscleral explantation in 2008 led to a briefly increased contamination rate. CONCLUSION: Donor demographic data run parallel to the general demographic development. Our analysis indicates a dynamic development of the eye bank over the last 30 years and emphasizes the need for an active quality management in coping with the challenges of modern eye banking.


Assuntos
Córnea , Transplante de Córnea , Bancos de Olhos/estatística & dados numéricos , Preservação de Órgãos/tendências , Doadores de Tecidos/estatística & dados numéricos , Obtenção de Tecidos e Órgãos/tendências , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Causas de Morte , Contagem de Células , Bases de Dados Factuais , Feminino , Seguimentos , Alemanha/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Cultura de Órgãos , Estudos Retrospectivos , Adulto Jovem
19.
Rev. bras. oftalmol ; 73(6): 351-357, Nov-Dec/2014. tab, graf
Artigo em Português | LILACS | ID: lil-741916

RESUMO

Objetivo: Traçar o perfil das doações de córnea obtidas pelo Banco de Tecidos Oculares do Hospital Getúlio Vargas (BTOC-HGV), avaliar a qualidade do tecido captado e enumerar as indicações do transplante de córnea. Métodos: Estudo retrospectivo de ficha de doadores cadastrados no BTOC-HGV no período de 2008 a 2011. Foram coletados dados de idade, gênero e causa do óbito do doador, tempo de enucleação, tempo de preservação, qualidade da córnea doada, causas de descartes e indicações de transplante de córnea no BTOC. Resultados: Foram analisadas 311 fichas de doadores de córnea do BTOC-HGV. O número de doações aumentou de 9 em 2008 para 80 em 2009, 109 em 2010 e 113 em 2011. A maioria dos doadores era do gênero masculino. A média da idade dos doadores foi de 43,00 ± 16,01 anos. A maioria das córneas transplantadas (78,6%) foi óptica. Foram descartadas 7,16% das córneas ópticas, sendo a maioria por nova classificação em tectônica e detecção de sorologia positiva. O tempo de enucleação foi de 3,77 ± 2,18 horas e o de preservação foi de 6,86 ± 6,10 horas. A causa mortis mais frequente foram causas externas, seguida por doenças do aparelho circulatório, respiratório, digestivo e geniturinário. A principal indicação de transplante foi ceratopatia bolhosa, seguida por ceratocone e perfuração corneana. Conclusão: Este estudo demonstrou a existência de fatores associados à qualidade das córneas captadas pelo BTOC como idade, tempo entre a enucleação e a preservação e causa do óbito. A principal indicação de transplante foi ceratopatia bolhosa. .


Objective: To describe the profile of corneal donations obtained by Ocular Tissue Bank, Hospital Getúlio Vargas (BTOC-HGV), assess the quality of the captured tissue and list the indications for corneal transplant. Methods: Retrospective study of registered donors in BTOC-HGV in the period 2008-2011. Collected data were: age, gender and cause of death of the donor, enucleation time, preservation time, quality of the donated cornea, causes of discards and indications for penetrating keratoplasty in BTOC. Results: Were analyzed 311 records of donor cornea BTOC-HGV. The number of donations increased from 9 in 2008 to 80 in 2009, 109 in 2010 and 113 in 2011. Most donors were male. The average age of the donors was 43.00 ± 16.01 years. The majority of transplanted corneas (78.6%) was optics. Were discarded 7.16% of corneal optics, mostly by new tectonic classification and detection of positive serology. The time of enucleation was 3.77 ± 2.18 hours and preservation was 6.86 ± 6.10 hours. The most frequent causes of death were external causes, followed by diseases of the circulatory, respiratory, digestive and genitourinary systems. The main indication for transplantation was bullous keratopathy, followed by keratoconus and corneal perforation. Conclusion: This study demonstrated the existence of factors associated with quality of corneas by BTOC as age, time between enucleation and preservation and cause of death. The main indication for transplantation was bullous keratopathy. .


Objective: To describe the profile of corneal donations obtained by Ocular Tissue Bank, Hospital Getúlio Vargas (BTOC-HGV), assess the quality of the captured tissue and list the indications for corneal transplant. Methods: Retrospective study of registered donors in BTOC-HGV in the period 2008-2011. Collected data were: age, gender and cause of death of the donor, enucleation time, preservation time, quality of the donated cornea, causes of discards and indications for penetrating keratoplasty in BTOC. Results: Were analyzed 311 records of donor cornea BTOC-HGV. The number of donations increased from 9 in 2008 to 80 in 2009, 109 in 2010 and 113 in 2011. Most donors were male. The average age of the donors was 43.00 ± 16.01 years. The majority of transplanted corneas (78.6%) was optics. Were discarded 7.16% of corneal optics, mostly by new tectonic classification and detection of positive serology. The time of enucleation was 3.77 ± 2.18 hours and preservation was 6.86 ± 6.10 hours. The most frequent causes of death were external causes, followed by diseases of the circulatory, respiratory, digestive and genitourinary systems. The main indication for transplantation was bullous keratopathy, followed by keratoconus and corneal perforation. Conclusion: This study demonstrated the existence of factors associated with quality of corneas by BTOC as age, time between enucleation and preservation and cause of death. The main indication for transplantation was bullous keratopathy. .


Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Doadores de Tecidos/estatística & dados numéricos , Obtenção de Tecidos e Órgãos/estatística & dados numéricos , Transplante de Córnea/estatística & dados numéricos , Coleta de Tecidos e Órgãos/estatística & dados numéricos , Seleção do Doador/estatística & dados numéricos , Bancos de Olhos/estatística & dados numéricos , Preservação de Órgãos , Fatores de Tempo , Causas de Morte , Córnea/patologia
20.
São Paulo; s.n; 2014. 63 p. ilus, tab, graf.
Tese em Português | LILACS | ID: lil-750112

RESUMO

Introdução: Doador de rim em vida é uma importante fonte de órgão para os pacientes portadores de doença renal crônica (DRC). Os doadores experimentam uma redução abrupta da taxa de filtração glomerular (TFG) e adaptações ao metabolismo mineral demandam estudos nesta população. Nós avaliamos prospectivamente esta adaptação em doadores de rim em vida. Métodos: Entre janeiro de 2010 a agosto de 2011, no hospital das Clínicas de São Paulo e na Universidade de Miami, realizamos a avaliação prospectiva do metabolismo mineral e da função renal por 1 ano em 74 doadores de rim em vida. Medimos a taxa de filtração glomerular (TFG), fósforo (Pi), cálcio (Ca), paratohormônio (PTH), fibroblast Growth Factor 23 (FGF23) e a fração de excreção do fósforo (FePO4) no pré-operatório e nos dias 1, 2, 14, 180 e 360 do pós-operatório. Resultados: Observamos uma redução, aproximadamente, de 40% da TFG nos dois primeiros dias após a cirurgia. No décimo quarto dia após a nefrectomia, observamos o início da recuperação da TFG, chegando ao máximo da recuperação com 1 ano, quando se atingiu 68,6% da função renal se comparado com o dia anterior a doação (75,3 ml/min/1,73m2, p < 0,001). O cálcio sérico apresentou seu nadir no dia 1 (7,99 mg/dL; p < 0,01) e o Pi sérico atingiu seu nadir no dia 2 (2,61 mg/dL; p < 0,01). Já no dia 14, os valores de Ca e Pi retornaram aos valores basais tendo o fósforo evoluído novamente com valores inferiores ao basal no último dia de seguimento (3,36mg/dL; p < 0,001). FGF23 e PTH apresentaram elevação no D1 (111,0144,6 percentil 25-75: 16-63 RU/ml 64,9 30,3pg/mL; p < 0,01). Os valores de FGF23 se mantiveram elevados até o final do estudo enquanto que o PTH retornou aos valores de base no segundo dia e, a partir de então, manteve sem diferença do valores basais até o último dia de estudo. FePO4 elevou de 11,45,2% para 15,28,1% entre o pré-operatório e D365 (p < 0,01). Conclusão: A nefrectomia para doação de rim em 74 pacientes saudáveis elevou os valores...


Introduction: Living kidney donors (LKDs) experience an abrupt decline in glomerular filtration rate (GFR). Mineral metabolism adaptations in early CKD are still debated and not well studied in LKDs. We prospectively studied acute and long term mineral metabolism adaptation of LKDs. Materials and Methods: We measured renal function and mineral metabolites longitudinally for 1 year (days (D) 1, 2, 14, 180, & 365 post-operatively) in 74 healthy individuals who underwent kidney live donation. Results: eGFR (MDRD) decreased to 59% of its baseline on day 2 and started to increase at day 3, to its maximum at day 360 (75.3±15.6 ml/min/1.73m2, p < 0.01) wile FGF23 increased from 60.6 (25th-75th percentile 19-81 RU/mL) at baseline to 111.0±144.6 (p < 0.01) on day 1 and keep higher than baseline throwout the study. PTH rose maximally on day 1 (64.9 ± 30.3pg/ml) and returned to its base line on D2 and did not change after that. Total serum Calcium levels decreased from 9,40±0,48 mg/dL to a nadir of 7.99±0,51 mg/dL on day 1 (p < 0.001). Serum Phosphate levels reached their nadir on day 2 (2.61±0,52 mg/dL; p < 0.01). At D14 total calcium and phosphate levels had returned to baseline, but phosphate levels returned down on D360 (3.36±0,52 mg/dL; p < 0.001). Phosphate excretion fraction (FePO4) increased from base line (11.4±5.2%) up to 15.2±8.1% until D360 (p < 0.001). Conclusions: Abrupt reduction in eGFR induces physiological increases in FGF23 and PTH, and decreases in serum Ca and Pi in the first week. The changes in FGF23 and Pi urinary fractional excretion of Pi remain modestly yet significantly different from baseline throughout the first year after nephrectomy. Wile Ca, PTH and Pi serum levels are not significantly different from the baseline...


Assuntos
Humanos , Adolescente , Adulto Jovem , Pessoa de Meia-Idade , Fatores de Crescimento de Fibroblastos , Transplante de Rim , Doadores Vivos , Minerais/metabolismo , Nefrectomia , Hormônio Paratireóideo , Doadores de Tecidos
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