- Publikační typ
- abstrakt z konference MeSH
Hyaluronic acid (HA) is an acidic, non-sulfated glycosaminoglycan that is intensively studied as a biodegradable and biocompatible material for scaffolding, regenerative medicine, and clinical applications [...].
- MeSH
- biokompatibilní materiály * terapeutické užití MeSH
- kyselina hyaluronová * MeSH
- lidé MeSH
- regenerativní lékařství MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- úvodníky MeSH
Despite several scientific or ethical issues, fetal bovine serum (FBS) remains the standard nutrient supplement in the mesenchymal stem cell cultivation medium. Cell amplification plays an important role in human stem cell therapies. Increasing interest in this field has supported attempts to find suitable human alternatives to FBS for in vitro cell propagation. Human platelet lysate (hPL) has recently been determined as one of them. Our study aimed to evaluate the influence of 2% hPL in the growth medium for in vitro expansion of human natal dental pulp stem cells (hNDP-SCs). The effect was determined on proliferation rate, viability, phenotype profile, expression of several markers, relative telomere length change, and differentiation potential of four lineages of hNDP-SCs. As a control, hNDP-SCs were simultaneously cultivated in 2% FBS. hNDP-SCs cultivated in hPL showed a statistically significantly higher proliferation rate in initial passages. We did not observe a statistically significant effect on mesenchymal stem cell marker (CD29, CD44, CD73, CD90) or stromal-associated marker (CD13, CD166) expression. The cell viability, relative telomere length, or multipotency remained unaffected in hNDP-SCs cultivated in hPL-medium. In conclusion, hPL produced under controlled and standardized conditions is an efficient serum supplement for in vitro expansion of hNDP-SCs.
- MeSH
- buněčná diferenciace MeSH
- kultivované buňky MeSH
- lidé MeSH
- mezenchymální kmenové buňky * MeSH
- proliferace buněk MeSH
- živiny MeSH
- zubní dřeň * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The physical stresses during cryopreservation affect stem cell survival and further proliferation. To minimize or prevent cryoinjury, cryoprotective agents (CPAs) are indispensable. Despite the widespread use of 10% dimethyl sulfoxide (DMSO), there are concerns about its potential adverse effects. To bypass those effects, combinations of CPAs have been investigated. This study aimed to verify whether high-molecular-hyaluronic acid (HMW-HA) serves as a cryoprotectant when preserving human mesenchymal stem cells (hMSCs) to reduce the DMSO concentration in the cryopreservation medium. We studied how 0.1% or 0.2% HMW-HA combined with reduced DMSO concentrations (from 10% to 5%, and 3%) affected total cell count, viability, immunophenotype, and differentiation potential post-cryopreservation. Immediately after cell revival, the highest total cell count was observed in 10% DMSO-stored hMSC. However, two weeks after cell cultivation an increased cell count was seen in the HMW-HA-stored groups along with a continued increase in hMSCs stored using 3% DMSO and 0.1% HMW-HA. The increased total cell count corresponded to elevated expression of stemness marker CD49f. The HA-supplemented cryomedium did not affect the differential potential of hMSC. Our results will participate in producing a ready-to-use product for cryopreservation of mesenchymal stem cells.
This work describes and evaluates vaccination against COVID-19 among members of the Czech Dental Chamber during the pre-booster vaccination phase. A cross-sectional online survey was conducted between 23 June and 4 September 2021, among 2716 participants, representing 24.3% of all chamber members. A total of 89.5% of respondents stated that they were registered for vaccination against COVID-19, their vaccination had started or been completed, or had a medically relevant reason to avoid vaccination. A total of 79.6% of respondents stated that they were fully vaccinated, most of them with the Comirnaty (Pfizer-BioNTech) vaccine (88.3%). The vaccination rate among males was significantly higher than among females (p = 0.001, OR 1.48). The main reasons for vaccination were professional (91.5%). The share of fully vaccinated participants was significantly higher (p < 0.0001, OR = 8.17) compared to the Czech general population (30.8%). A COVID-19 vaccine breakthrough infection rate was 0.42%. The study shows that both the willingness to vaccinate and the proportion of fully vaccinated individuals among Czech dentists are high, and only about 10% of them refused vaccination based on reasons classified as not medically relevant.
- Publikační typ
- časopisecké články MeSH
It is primarily important to define the standard features and factors that affect dental pulp stem cells (DPSCs) for their broader use in tissue engineering. This study aimed to verify whether DPSCs isolated from various teeth extracted from the same donor exhibit intra-individual variability and what the consequences are for their differentiation potential. The heterogeneity determination was based on studying the proliferative capacity, viability, expression of phenotypic markers, and relative length of telomere chromosomes. The study included 14 teeth (6 molars and 8 premolars) from six different individuals ages 12 to 16. We did not observe any significant intra-individual variability in DPSC size, proliferation rate, viability, or relative telomere length change within lineages isolated from different teeth but the same donor. The minor non-significant variances in phenotype were probably mainly because DPSC cell lines comprised heterogeneous groups of undifferentiated cells independent of the donor. The other variances were seen in DPSC lineages isolated from the same donor, but the teeth were in different stages of root development. We also did not observe any changes in the ability of cells to differentiate into mature cell lines-chondrocytes, osteocytes, and adipocytes. This study is the first to analyze the heterogeneity of DPSC dependent on a donor.
- MeSH
- buněčná diferenciace fyziologie MeSH
- buněčné linie MeSH
- buněčný rodokmen fyziologie MeSH
- chondrocyty fyziologie MeSH
- dárci tkání MeSH
- individuální biologická variabilita MeSH
- kmenové buňky fyziologie MeSH
- lidé MeSH
- mladiství MeSH
- osteocyty fyziologie MeSH
- proliferace buněk fyziologie MeSH
- tukové buňky fyziologie MeSH
- zubní dřeň fyziologie MeSH
- Check Tag
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Alveolární ostitida neboli suché lůžko je častou postextrakční komplikací. Klinicky nacházíme částečně až zcela dezintegrované koagulum v extrakční ráně, výrazný foetor ex ore a značnou bolest lokalizovanou v lůžku po extrahovaném zubu, někdy s iradiací do ucha a spánkové krajiny na postižené straně. Cílem této studie bylo zhodnotit výsledek léčby alveolární ostitidy nově formulovaným lokálním preparátem obsahujícím kyselinu hyaluronovou a oktenidin dihydrochlorid. Tato antisepticky působící směs ulpívá na sliznici a vyplňuje zubní lůžko, čímž poskytuje obnažené kosti v extrakční ráně mechanickou ochranu. Do studie bylo zahrnuto celkem 49 pacientů s diagnózou alveolární ostitidy. Testovaný přípravek byl opakovaně aplikován 1x denně, a to do ústupu lokální bolesti na méně než 20 mm vizuální analogové stupnice. Pokud bylo dosaženo těchto hodnot do 7 dní, byla léčba považována za efektivní. Čtvrtý den po zahájení studie byla úspěšnost léčby 71,7 %, sedmý den činila již 87 %. Intenzita bolesti v době mezi prvním vyšetřením a sedmou aplikací poklesla o 95 % při hodnocení osmého dne. Tento rozdíl byl statisticky významný (p < 0,0001). Získaná data potvrdila, že klinicky testovaný prostředek s obsahem lyofilizované kyseliny hyaluronové a oktenidinu je v terapii alveolární ostitidy dostatečně efektivní. Během popsané léčby nebyly pozorovány žádné vedlejší ani nežádoucí účinky.
Alveolar osteitis is a complication associated relatively often with the extraction of a tooth. Clinically, there is usually found a socket without blood cloth, halitosis and intensive localized pain, sometimes with the irradiation towards the ipsilateral ear and temporal region. The aim of this study was to assess the outcome of the treatment of alveolar osteitis using a new medical compound consisted of the hyaluronic acid and octenidine dihydrochloride. This fully soluble and degradable material was designed to adhere well to the mucosa, obturate the empty socket, and disinfect it. A total amount of 49 patients were included in this study. The tested medical compound was administered repeatedly once a day until the local pain subsides on less than 20 mm of visual analog scale. Treatment was considered successful when the pain subsided to less than 20 mm of the scale during 7 days of treatment, not longer. The results of the study demonstrated 71.7% success rate after 4 days of treatment and 87% success rate after 7 days of treatment. The decrease of the pain between the initial examination and the 7th administration was 95%. This decrease was statistically significant (p <0.0001). The study confirmed the effectiveness of the lyophilized hyaluronic acid combined with octenidine dihydrochloride in the treatment of alveolar osteititis. No side effects of the therapy have been observed.
AIM: The aim of this study was to assess the perception of undergraduate dentistry students at Charles University, Faculty of Medicine in Hradec Králové, the Czech Republic regarding their endodontic education within the context of the Undergraduate Curriculum Guidelines for Endodontology by the European Society of Endodontology (ESE). The secondary aim was to compare this perception among students in the Czech and English groups. METHODOLOGY: A questionnaire survey was conducted among fifth year students at the very end of their studies. RESULTS: The students returned 60 filled questionnaires, making the response rate of 75.9%. More than two thirds of the respondents declared that they were competent at or had knowledge of most of the major competencies defined by the ESE. Eighty seven percent of respondents felt competent to perform a root canal treatment on anterior teeth; 86.7% on premolars; and 48.3% on molars. Nearly all respondents (98.3%) recommended more opportunities to practice on patients. CONCLUSIONS: The overall perception of the students was that their endodontic education was sufficient and largely conformed to the guidelines. Insufficient exposure to endodontic practice on patients was identified as a deficiency. There were no significant differences in perceptions between the two study groups.
- MeSH
- dospělí MeSH
- endodoncie výchova MeSH
- jazyk (prostředek komunikace) MeSH
- klinické kompetence * MeSH
- kurikulum MeSH
- lidé MeSH
- mladý dospělý MeSH
- percepce MeSH
- průzkumy a dotazníky MeSH
- sebeuplatnění * MeSH
- studenti stomatologie * psychologie MeSH
- studium stomatologie normy MeSH
- terapie kořenového kanálku MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Úvod: Zubní dřeň představuje relativně snadno dostupný zdroj mezenchymových kmenových buněk. Kmenové buňky ze zubní dřeně (KBZD) byly poprvé izolovány v roce 2000 a od té doby jsou intenzivně studovány. Cíl: Cílem studie bylo optimalizovat proces izolace KBZD, zejména zkrátit dobu, po kterou je zubní dřeň enzymaticky štěpena. U KBZD izolovaných novým postupem stanovit proliferační aktivitu, fenotyp, viabilitu a ověřit schopnost jejich diferenciace ve zralé buněčné typy – osteoblasty, chondroblasty a adipocyty. Materiály a metodika: Extrahovali jsme celkem pět stálých zubů a izolovali pět buněčných linií, které jsme kultivovali v modifikovaném kultivačním médiu (α-MEM) pro adultní mezenchymové progenitorové buňky, obohaceném o 2 % fetálního bovinního séra, růstové faktory, antibiotika, antimykotika, a doplněném o Insulin-Transferrin-Sodium-Selenium supplement. Pro stanovení viability, počtu a velikosti buněk jsme použili přístroje Vi-Cell analyzer a Z2-Counter. Fenotypová analýza byla provedena pomocí průtokového cytometru Cell Lab Quanta. Pro diferenciaci v chondroblasty, osteoblasty a adipocyty jsme použili komerčně dostupná diferenciační média. Průkaz diferenciace jsme dokazovali imunohistochemicky (osteokalcin a kolagen typu II) a histologickým barvením (modrý Massonův trichrom, barvení dle Kossy a olejová červeň). Výsledky: KBZD jsme kultivovali do 8. pasáže. Buňky dosáhly průměrně 47,8 ± 2,0 populačních zdvojení (angl. population doublings; PD). Průměrný čas potřebný pro zdvojení populace (angl. doubling time; DT), činil 39,2 ± 6,1 hodin. Po celou dobu kultivace byly buňky proliferačně aktivní. Průměrná viabilita v 2. pasáži byla 92,3 ± 1,5 % a v 8. pasáži 92,4 ± 1,4 %. Fenotypovou analýzou jsme prokázali vysokou expresi povrchových antigenů pro mezenchymové kmenové buňky (angl. cluster of differentiation; CD) CD13, CD29, CD44, CD90, pro tzv. „stromal associated“ znaky CD73, CD166, a naopak nízkou či nízce pozitivní expresi znaků CD31, CD34, CD45, typických pro endoteliální a hematopoetickou buněčnou řadu. KBZD se diferencovaly ve zralé buněčné typy, osteoblasty a chondroblasty. I přes proadipogenně silně působící médium buňky nediferencovaly v adipocyty. Závěr: Podařilo se nám optimalizovat izolační protokol pro KBZD tím, že jsme zkrátili dobu, po kterou je zubní dřeň enzymaticky štěpena. KBZD izolované touto metodou prokazovaly po celou dobu kultivace vysoký proliferační a diferenciační potenciál. Nezpozorovali jsme žádné známky spontánní diferenciace či degenerace. Právě pro vysokou proliferační aktivitu, široký diferenciační potenciál a snadnou dostupnost představují KBZD budoucnost v regenerativní medicíně.
Introduction: The dental pulp represents an easily accessible source of adult dental pulp stem cells (DPSCs). They were isolated for the first time in 2000 and since then many researchers have investigated and analysed their biological characteristics. Aim: The purpose of this study was to optimize the isolation protocol for DPSCs, namely to shorten the time of enzymatic digestion of the dental pulp, and to cultivate isolated DPSCs using this new approach, investigate their proliferation, phenotype, cell viability and determine their ability to differentiate into mature cells, chondroblasts, osteoblasts, and adipocytes. Materials and methods: Out of five extracted permanent teeth, we isolated five dental pulp stem cell lineages. They were cultivated in a modified cultivation media (α-MEM) for mesenchymal adult progenitor cells containing 2 % fetal bovine serum (FBS) and supplemented with growth factors, antibiotics, antimycotics and Insulin-Transferrin-Sodium-Selenium supplement (ITS). The cell viability, cell count and other properties were examined using a Vi-Cell analyzer and Z2-Counter. The phenotype analysis was performed using a flow cytometer Cell Lab Quanta. For differentiation in chondroblasts, osteoblasts and adipocytes, we used commercially available differentiation media. The evidence of differentiation was proved by the immunocytochemistry (osteocalcin and collagen type II) and histological staining (blue Masson's trichrome, von Koss stain and oil red). Results: We were able to cultivate DPSCs over 47.8 ± 2.0 population doublings (PD). The average population doubling time (DT) was 39.2 ± 6.1 hours. The average cell viability was 92.3 ± 1.5 % in the second passage and 92.4 ± 1.4 % in the eighth passage. DPSCs showed high positivity for mesenchymal stem cell markers (cluster of differentiation; CD) CD29, CD44, CD90 and for stromal associated markers CD13, CD73, CD166 and negative expression or low positivity for hematopoietic markers CD34, CD45 and for CD31. DPSCs differentiated into osteoblasts and chondroblasts. Even after the exposition of the strong adipogenic medium they did not show any signs of differentiation into adipocytes. Conclusion: We have successfully optimized the isolation protocol for DPSCs by shortening the time of enzymatic digestion of the dental pulp. DPSCs isolated using the new method demonstrated the high proliferation and differentiation potential throughout long-term cultivation. We did not observe any signs of spontaneous differentiation or cell degeneration. DPSCs seems to be the promising future for a regenerative and reparative medicine thanks to their remarkable high proliferative potential and ability to differentiate into many mature cell populations.
Úvod a cíl práce: Kmenové buňky zubní dřeně (KBZD) vykazují přirozeně vysokou pozitivitu na povrchový receptor glykoprotein CD44 podílející se mimo jiné na indukci mineralizace odontoblastů, přičemž kyselina hyaluronová (KH) je jeho hlavní ligandou. Cílem experimentu bylo posoudit vliv KH o nízké (NMH-KH), střední (SHM-KH) a vysoké (VMH-KH) molekulární hmotnosti na fenotypový profil, proliferační aktivitu a diferenciační potenciál lidských KBZD. Metodika: Experiment byl proveden v in vitro podmínkách na dvou liniích lidských KBZD odlišných dárců (třetí molár – muž, 25 let, a první premolár – muž, 9 let). Tyto linie byly kultivovány ve standardním médiu a od druhé pasáže také ve třech experimentálních kultivačních médiích obsahujících 0,1 % kyseliny hyaluronové (KH) ve třech molekulárních hmotnostech: NMH-KH (116 kDa), SHM-KH (540 kDa) a VHM-KH (1500 kDa). Analýza fenotypu byla provedena v sedmé pasáži pomocí průtokového cytometru Vi-Cell XR, viabilita byla měřena analyzátorem Vi-Cell v sedmé pasáži a počet buněk analyzátorem Z2 Coulter ve všech pasážích. Osteodiferenciace a chondrodiferenciace byly indukovány komerčně dodávanými diferenciačními médii a prokazovány histologickým barvením s alciánovou modří a alizarinovou červení. Výsledky: Linie KBZD využité v našem experimentu vykazovaly fenotyp typický pro lidské KBZD (vysoká pozitivita pro CD 13, CD 29, CD 44, CD 90 a OCT3/4), byly schopny překročit Hayflickův limit a diferencovat v buňky produkující osteogenní a chondrogenní extracelulární matrix. Během experimentu dosáhly KBZD linie 1 kultivované v kontrolním médiu / médiu 1 (116 kDa KH) / médiu 2 (540 kDa KH) / médiu 3 (1500 kDa KH) v tomto pořadí celkem 14,1 / 15,3 / 15,4 / 14,8 populačních zdvojení. Medián doubling time s minimální a maximální hodnotou ve stejném pořadí byl 31,6 (29,2; 36,8) / 29,6 (28,5; 30,6) / 30,3 (26,7; 31,3) / 30,5 (29,7; 33,8) hodin. Viabilita KBZD získaných ze sedmé pasáže byla ve stejném pořadí 92,3 / 93,1 / 91,8 / 92,6 %. KBZD linie 2 kultivované v kontrolním médiu / médiu 1 (116 kDa KH) / médiu 2 (540 kDa KH) / médiu 3 (1500 kDa KH) dosáhly v tomto pořadí celkem 16,7 / 17,2 / 16,7 / 16,8 populačních zdvojení. Medián doubling time s minimální a maximální hodnotou ve stejném pořadí byl 28,9 (24,5; 35,2) / 27,4 (24,5; 32,6) / 28,3 (24,0; 38,4) / 27,1 (25,3; 33,9) hodin. Viabilita KBZD získaných ze sedmé pasáže byla ve stejném pořadí 80,1 / 82,5 / 81,8 / 80,9 %. Závěr: Ověřili jsme, že KBZD v přítomnosti kyseliny hyaluronové o molekulárních hmotnostech 116, 540 a 1500 kDa v koncentraci 0,1 % přežívají, proliferují a udržují si schopnost diferencovat ve zralé buněčné elementy. Dále jsme ověřili původní předpoklad, že nízkomolekulární forma kyseliny hyaluronové bude mít rozdílný dopad na fenotyp KBZD než forma vysokomolekulární.
Introduction: Dental pulp stem cells (DPSCs) express naturally high positivity for surface receptor glycoprotein CD 44 which is involved in the induction of odontoblast mineralization with hyaluronic acid (HA) being its major ligand. The aim of this experiment was to assess the effect of HA in low (LMW-HA), medium (MMW-HA) and high (HMW-HA) molecular weights on the phenotypic profile, proliferation activity and differentiation potential of human DPSCs. Methods: The experiment was conducted in vitro on two lines of human DPSCs from different donors (third molar – male, 25 years and first premolar – male, 9 years). These lines were cultured in standard medium and from the second passage also in three experimental culture media containing 0.1% HA in three molecular weights: LMW-HA (116 kDa), MMW-HA (540 kDa) and HMW-HA (1500 kDa). The phenotypic analysis was performed in the seventh passage using a Vi-Cell XR flow cytometer, viability was evaluated by the Vi-Cell Analyzer in the seventh passage and proliferation activity measured by the Z2 Counter Analyzer in every passage. Osteo- and chondro- differentiation were inducted by commercially supplied cultivation media and demonstrated by histological staining with alcian blue and alizarin red. Results: DPSCs used in our experiment expressed phenotype typical for human DPSCs (high positivity for CD 13, CD 29, CD 44, CD 90 and OCT 3/4), they were able to exceed Hayflick limit and differentiate in the osteogenic as well as the chondrogenic extracellular matrix. During the experiment, DPSCs line 1 cultivated in control medium / medium 1 (116 kDa HA) / medium 2 (540 kDa HA) / medium 3 (1500 kDa HA) achieved in this order in total 14.1/15.3/15.4/14.8 population doublings. The median of doubling time with the minimal and maximal values in the same order was 31.6 (29.2; 36.8) / 29.6 (28.5; 30.6) / 30.3 (26.7; 31.3) / 30.5 (29.7; 33.8) hours. The viability of the DPSCs obtained from the seventh passage was in the same order 92.3/93.1/91.8/92.6%. DPSC line 2 cultivated in control medium / medium 1 (116 kDa HA) / medium 2 (540 kDa HA) / medium 3 (1500 kDa HA) achieved in this order in total 16.7/17.2/16.7/16.8 population doublings. The median of doubling time with the minimal and maximal values in the same order was 28.9 (24.5; 35.2) / 27.4 (24.5; 32.6) / 28.3 (24.0; 38.4) / 27.1 (25.3; 33.9) hours. The viability of the DPSCs obtained from the seventh passage was in the same order 80.1/82.5/81.8/80.9%. Conclusion: We verified that DPSCs in the presence of hyaluronic acid at a concentration of 0.1% and molecular weights of 116, 540 and 1500 kDa survive, proliferate and maintain the ability to differentiate in mature cellular elements. We also verified the original assumption that the low molecular weight form of hyaluronic acid has a different impact on the DPSCs‘ phenotype than the high molecular weight form of hyaluronic acid.
- MeSH
- kyselina hyaluronová * MeSH
- lidé MeSH
- výzkum kmenových buněk * MeSH
- zubní dřeň MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
