Cíl: Představit použití lidské kryokonzervované amniové membrány (AM) v léčbě chronických ran, specifikovat indikační kritéria pro její aplikaci, protokol aplikace a metodiku převazů. Dalším cílem je seznámení se souhrnnými výsledky studie léčby chronických nehojících se ran pomocí AM a s predikcí zhojení dle dynamiky léčby. Materiály a metody: Standardizace léčby pomocí kryokonzervované AM proběhla v rámci multicentrické studie ve třech klinických centrech. Pacientům s chronickými ranami, kteří se nehojili v průměru po dobu 34 měsíců (minimálně šest týdnů), byla aplikována AM, s pravidelnými vizitami po sedmi dnech. Výsledky: Z 35 pacientů s 43 ranami různé etiologie, bylo po aplikaci AM zcela zhojeno 24 ran (56 %), 10 ran bylo zhojeno částečně (> 50 % plochy rány) (23 %), 9 ran (21 %) se zhojit nepodařilo. Během léčby AM zaznamenali pacienti silný analgetický účinek. Sledování dynamiky hojení vedlo ke stanovení predikčního intervalu pro posouzení schopnosti rány zhojit se pomocí AM na 10–12 týdnů. Závěr: Léčba chronických ran pomocí AM přináší prospěch indikovaným pacientům, jak v akceleraci hojení, tak ve zlepšení kvality života, a to zejména snížení bolesti v oblasti rány.
Objective: This work presents the use of human cryopreserved amniotic membrane (AM) in the treatment of chronic wounds, specifying the indication criteria for its application, application protocol, and wound dressing methodology. At the same time, it presents the summary results of the study of the treatment of chronic non-healing wounds using AM with the prediction of healing according to the dynamics of the treatment. Materials and methods: Standardization of treatment using cryopreserved AM took place within the framework of a multicentre study in three specialist centres. Patients with chronic wounds that did not heal for an average of 34 months (min. six weeks) were treated with AM, with regular visits every seven days. The AM application was carried out 1–2 times in 14 days. Results: Out of 35 patients with 43 wounds of various etiologies, 24 wounds (56%) were completely healed after AM application, 10 wounds were partially healed (> 50% of the wound area) (23%), 9 wounds (21%) failed to heal. During AM treatment, patients experienced a strong analgesic effect. Monitoring the dynamics of healing led to the establishment of a prediction interval for assessing the ability of the wound to heal using AM at 10–12 weeks. Conclusion: The treatment of chronic wounds using AM brings benefits to the indicated patients, both in the acceleration of healing and in the improvement of the quality of life, especially in the reduction of pain in the wound area.
- MeSH
- amnion * fyziologie MeSH
- biologické krytí * klasifikace MeSH
- hojení ran MeSH
- kryoprezervace metody MeSH
- lidé MeSH
- membrány fyziologie MeSH
- placenta fyziologie MeSH
- rány a poranění ošetřování terapie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- multicentrická studie MeSH
- práce podpořená grantem MeSH
Autologous serum eye drops (ASEDs) are used as a treatment for severe dry eye disease. The concentration and stability of various growth factors in ASEDs is determinative for their efficiency. We therefore assessed the concentrations of transforming growth factor beta 1 (TGF-β1), epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF-1) in ASEDs following storage at 4-8, -20, -80 and -156 °C. Twenty % and 100% sera from eight healthy volunteers were analysed by the sandwich enzyme immunoassay at different time intervals up to seven months. The mean levels of TGF-β1 and EGF in undiluted and 20% serum did not differ significantly from the baseline levels in fresh serum for any storage conditions after 7 days at 4-8 °C, as well as after 4- and 7-month preservation at sub-zero temperatures. In 20% serum, no IGF-1 concentration decrease was found following 7 days of preservation at 4-8 °C. However, a decrease to 78 % and 81 % (P < 0.01) of baseline values was found in 20% serum after 4-month storage at -20 °C and 7-month storage at -156 °C, respectively. A more pronounced decrease in IGF-1 was observed in undiluted serum. All assessed growth factors present in 20% frozen serum remained stable for up to 7 months. The highest stability was achieved at -80 °C. At -20 and -156 °C, some decrease in IGF-1 occurred. Our results indicate that 20% ASEDs can be stored frozen up to 7 months under proper conditions.
- Publikační typ
- abstrakt z konference MeSH
Yeast cells in general are known to be difficult to prepare for electron microscopy investigations particularly when the preservation of antigenicity is required. In this work, we compare various protocols for preparation of Saccharomyces cerevisiae cells for immunoelectron microscopy, ranging from classical chemical fixation to high-pressure freezing followed by freeze-substitution in different kinds of substitution media. Our aim was to establish a protocol giving optimal, routinely reproducible results for simultaneous retention of fine ultrastructural details and antigen immunoreactivity, with particular focus on the preservation of nuclear and nucleolar architecture. This was demonstrated by ultrastructural immunolocalization of various nucleolar (Nop1 and Nsr1), nuclear (Nsp1) and alpha-tubulin antigens. The protocol which we found to yield the best preserved Saccharomyces cerevisiae cells for both morphological and immunological studies included cryo-fixation by high-pressure freezing followed by freeze-substitution in acetone with 0.1% uranyl acetate and embedding in Lowicryl HM20. In addition, immunofluorescence detection of the antigens was performed and correlated with immunolabelling at the electron microscopy level.
- MeSH
- aceton chemie MeSH
- antigeny fungální chemie MeSH
- buněčné jadérko imunologie ultrastruktura MeSH
- buněčné jádro imunologie ultrastruktura MeSH
- financování organizované MeSH
- fluorescenční mikroskopie MeSH
- formaldehyd chemie MeSH
- glutaraldehyd chemie MeSH
- imunoelektronová mikroskopie metody MeSH
- mrazová substituce MeSH
- organokovové sloučeniny chemie MeSH
- Saccharomyces cerevisiae ultrastruktura MeSH
- transmisní elektronová mikroskopie MeSH
