Hypofosfatázie (HPP) je vzácné onemocnění způsobené nedostatečnou produkcí alkalické fosfatázy (ALP). Dochází k demineralizaci skeletu se závažnými extraskeletálními symptomy včetně postižení dentice, a to k předčasné ztrátě dočasných zubů ve frontálním úseku čelistí. Diagnostika je založena na nízké koncentraci ALP v séru v kombinaci s typickými klinickými projevy. Od roku 2015 je dostupná substituční enzymatická léčba pomocí rekombinantní ALP. X-vázaná hypofosfatemická křivice (XLH) je nejčastějším projevem křivice a osteomalacie. Projevuje se růstovou retardací, deformitami dolních končetin, bolestí svalů a kostí. Dentální manifestace se projevuje dysplazií dentinu s tvorbou abscesů u intaktních zubů, především v dočasné dentici. Léčba spočívá v pravidelné aplikaci protilátky proti FGF23 (fibroblastový růstový hormon 23), burosumabu. Burosumab je rekombinantní lidská monoklonální protilátka (IgG1), která se váže na FGF23 a tlumí jeho aktivitu.
Hypophosphatasia (HPP) is a rare disease caused by insufficient production of alkaline phosphatase. Main symptoms are skeletal demineralisation and severe extra-skeletal complications including dental symptoms. Dental manifestation is characterized by early loss of primary anterior teeth. Diagnosis is based on a low concentration of ALP in serum. Enzymatic substitution therapy with recombinant ALP is available since 2015. X-linked hypophosphatemia (XLH) is the most common form of rickets and osteomalacia. The disease manifests by growth retardation, deformities of lower limbs, bone and muscular pain. Dental manifestation is dentin dysplasia, followed by formation of periapical abscesses of intact teeth, mainly in primary dentition. A humanized monoclonal antibody for FGF23 (burosumab) is the promising treatment for XLH.
- MeSH
- abnormality zubů etiologie prevence a kontrola MeSH
- alkalická fosfatasa analýza genetika nedostatek terapeutické užití MeSH
- dítě MeSH
- familiární hypofosfatemická rachitida * diagnóza epidemiologie farmakoterapie genetika MeSH
- hypofosfatázie * diagnóza etiologie farmakoterapie MeSH
- lidé MeSH
- osteogeneze genetika MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- přehledy MeSH
Osteogenesis is an important process of bone metabolism, and abnormal osteogenesis leads to various skeletal system diseases. Osteoblasts, the main cells involved in bone formation, are central elements in the study of bone metabolic diseases. Single-cell RNA sequencing is an important tool for studying the transcriptome of cells and can help to elucidate various cellular and molecular functions at the single-cell level, providing new avenues for life science research. Here we explore the heterogeneity of osteoblasts and try to reveal the developmental trajectory of osteoblasts, thereby contributing to efforts to describe the mechanism of osteogenesis. In this study, single-cell sequencing data of murine bone marrow cells were used to identify osteoblasts. Finally, osteoblasts were divided into four groups, each differing in characteristic genes and signal pathways. We also identify clues of the changes of some genes in the process of osteoclast formation, providing directions for further study. Collectively, our findings suggest that bone marrow osteoblasts can be divided into several subgroups, which represent different stages of cells, and that the specific genes of each subgroup respond to the molecular mechanisms of cell development. This data will likely be of great help in resolving diseases of the skeletal system.
- MeSH
- buněčná diferenciace MeSH
- kostní dřeň * MeSH
- myši MeSH
- osteoblasty * metabolismus MeSH
- osteogeneze genetika MeSH
- osteoklasty metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Osteoblasts orchestrate bone formation through the secretion of type I collagen and other constituents of the matrix on which hydroxyapatite crystals mineralize. Here, we show that TENT5A, whose mutations were found in congenital bone disease osteogenesis imperfecta patients, is a cytoplasmic poly(A) polymerase playing a crucial role in regulating bone mineralization. Direct RNA sequencing revealed that TENT5A is induced during osteoblast differentiation and polyadenylates mRNAs encoding Col1α1, Col1α2, and other secreted proteins involved in osteogenesis, increasing their expression. We postulate that TENT5A, possibly together with its paralog TENT5C, is responsible for the wave of cytoplasmic polyadenylation of mRNAs encoding secreted proteins occurring during bone mineralization. Importantly, the Tent5a knockout (KO) mouse line displays bone fragility and skeletal hypomineralization phenotype resulting from quantitative and qualitative collagen defects. Thus, we report a biologically relevant posttranscriptional regulator of collagen production and, more generally, bone formation.
- MeSH
- buněčná diferenciace MeSH
- fyziologická kalcifikace genetika MeSH
- kolagen typu I, řetězec alfa 1 genetika metabolismus MeSH
- kolagen typu I genetika metabolismus MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši knockoutované MeSH
- myši MeSH
- neurotrofní faktory genetika metabolismus MeSH
- nukleotidyltransferasy genetika metabolismus MeSH
- oční proteiny genetika metabolismus MeSH
- osteoblasty metabolismus patologie MeSH
- osteogenesis imperfecta genetika metabolismus patologie MeSH
- osteogeneze genetika MeSH
- osteonektin genetika metabolismus MeSH
- poly(A)-polymerasa genetika metabolismus MeSH
- polyadenylace MeSH
- protein - isoformy nedostatek genetika MeSH
- sekvenční analýza RNA MeSH
- serpiny genetika metabolismus MeSH
- signální transdukce MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
MiR-21 is being gradually more and more recognized as a molecule regulating bone tissue homeostasis. However, its function is not fully understood due to the dual role of miR-21 on bone-forming and bone-resorbing cells. In this study, we investigated the impact of miR-21 inhibition on pre-osteoblastic cells differentiation and paracrine signaling towards pre-osteoclasts using indirect co-culture model of mouse pre-osteoblast (MC3T3) and pre-osteoclast (4B12) cell lines. The inhibition of miR-21 in MC3T3 cells (MC3T3inh21) modulated expression of genes encoding osteogenic markers including collagen type I (Coll-1), osteocalcin (Ocl), osteopontin (Opn), and runt-related transcription factor 2 (Runx-2). Inhibition of miR-21 in osteogenic cultures of MC3T3 also inflected the synthesis of OPN protein which is essential for proper mineralization of extracellular matrix (ECM) and anchoring osteoclasts to the bones. Furthermore, it was shown that in osteoblasts miR-21 regulates expression of factors that are vital for survival of pre-osteoclast, such as receptor activator of nuclear factor κB ligand (RANKL). The pre-osteoclast cultured with MC3T3inh21 cells was characterized by lowered expression of several markers associated with osteoclasts' differentiation, foremost tartrate-resistant acid phosphatase (Trap) but also receptor activator of nuclear factor-κB ligand (Rank), cathepsin K (Ctsk), carbonic anhydrase II (CaII), and matrix metalloproteinase (Mmp-9). Collectively, our data indicate that the inhibition of miR-21 in MC3T3 cells impairs the differentiation and ECM mineralization as well as influences paracrine signaling leading to decreased viability of pre-osteoclasts.
- MeSH
- buněčná diferenciace genetika MeSH
- buněčné linie MeSH
- extracelulární matrix metabolismus MeSH
- kokultivační techniky MeSH
- kyselá fosfatasa rezistentní k tartarátu metabolismus MeSH
- messenger RNA genetika MeSH
- mikro RNA genetika metabolismus MeSH
- myši MeSH
- osteoblasty metabolismus MeSH
- osteogeneze genetika MeSH
- osteoklasty metabolismus MeSH
- osteopontin genetika metabolismus MeSH
- parakrinní signalizace genetika MeSH
- protein PEBP2alfaA genetika metabolismus MeSH
- resorpce kosti metabolismus MeSH
- signální transdukce genetika MeSH
- transfekce MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tuftelin was originally discovered and mostly studied in the tooth, but later found also in other organs. Despite its wide distribution among tissues, tuftelin's function has so far been specified only in the formation of enamel crystals. Nevertheless, in many cases, tuftelin was suggested to be associated with cellular adaptation to hypoxia and recently even with cell differentiation. Therefore, we aimed to investigate tuftelin expression along with hypoxia-inducible factors (HIFs) during the early development of the mandibular/alveolar (m/a) bone, when osteoblasts started to differentiate in vivo and to compare their expression levels in undifferentiated versus differentiated osteoblastic cells in vitro. Immunohistochemistry demonstrated the presence of tuftelin already in osteoblastic precursors which were also HIF1-positive, but HIF2-negative. Nevertheless, HIF2 protein appeared when osteoblasts differentiated, one day later. This is in agreement with observations made with MC3T3-E1 cells, where there was no significant difference in tuftelin and Hif1 expression in undifferentiated vs. differentiated cells, although Hif2 increased upon differentiation induction. In differentiated osteoblasts of the m/a bone, all three proteins accumulated, first, prenatally, in the cytoplasm and later, particularly at postnatal stages, they displayed also peri/nuclear localization. Such a dynamic time-space pattern of tuftelin expression has recently been reported in neurons, which, as the m/a bone, differentiate under less hypoxic conditions as indicated also by a prevalent cytoplasmic expression of HIF1 in osteoblasts. However, unlike what was shown in cultured neurons, tuftelin does not seem to participate in final osteoblastic differentiation and its functions, thus, appears to be tissue specific.
- MeSH
- buňky 3T3 MeSH
- faktor 1 indukovatelný hypoxií analýza genetika MeSH
- imunohistochemie MeSH
- kultivované buňky MeSH
- myši MeSH
- osteogeneze genetika MeSH
- proteiny zubní skloviny analýza genetika MeSH
- transkripční faktory analýza genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Osteoblastic differentiation is a multistep process characterized by osteogenic induction of mesenchymal stem cells, which then differentiate into proliferative pre-osteoblasts that produce copious amounts of extracellular matrix, followed by stiffening of the extracellular matrix, and matrix mineralization by hydroxylapatite deposition. Although these processes have been well characterized biologically, a detailed transcriptional analysis of murine primary calvaria osteoblast differentiation based on RNA sequencing (RNA-seq) analyses has not previously been reported. Here, we used RNA-seq to obtain expression values of 29,148 genes at four time points as murine primary calvaria osteoblasts differentiate in vitro until onset of mineralization was clearly detectable by microscopic inspection. Expression of marker genes confirmed osteogenic differentiation. We explored differential expression of 1386 protein-coding genes using unsupervised clustering and GO analyses. 100 differentially expressed lncRNAs were investigated by co-expression with protein-coding genes that are localized within the same topologically associated domain. Additionally, we monitored expression of 237 genes that are silent or active at distinct time points and compared differential exon usage. Our data represent an in-depth profiling of murine primary calvaria osteoblast differentiation by RNA-seq and contribute to our understanding of genetic regulation of this key process in osteoblast biology.
- MeSH
- alternativní sestřih MeSH
- buněčná diferenciace genetika MeSH
- kultivované buňky MeSH
- lebka fyziologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- osteoblasty fyziologie MeSH
- osteogeneze genetika MeSH
- RNA analýza MeSH
- stanovení celkové genové exprese MeSH
- transkriptom genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The cellular and molecular basis of vascular calcification (VC) in atherosclerosis is not fully understood. Here, we investigate role of resident/circulating progenitor cells in VC and contribution of inflammatory plaque environment to this process. Vessel-derived stem/progenitor cells (VSCs) and mesenchymal stem cells (MSCs) isolated from atherosclerotic ApoE(-/-) mice showed significantly more in vitro osteogenesis and chondrogenesis than cells generated from control C57BL/6 mice. To assess their ability to form bone in vivo, cells were primed chondrogenically or cultured in control medium on collagen glycosaminoglycan scaffolds in vitro prior to subcutaneous implantation in ApoE(-/-) and C57BL/6 mice using a crossover study design. Atherosclerotic ApoE(-/-) MSCs and VSCs formed bone when implanted in C57BL/6 mice. In ApoE(-/-) mice, these cells generated more mature bone than C57BL/6 cells. The atherosclerotic in vivo environment alone promoted bone formation by implanted C57BL/6 cells. Un-primed C57BL/6 VSCs were unable to form bone in either mouse strain. Treatment of ApoE(-/-) VSC chondrogenic cultures with interleukin (IL)-6 resulted in significantly increased glycosaminoglycan deposition and expression of characteristic chondrogenic genes at 21 days. In conclusion, resident vascular cells from atherosclerotic environment respond to the inflammatory milieu and undergo calcification. IL-6 may have a role in aberrant differentiation of VSCs contributing to vascular calcification in atherosclerosis.
- MeSH
- apolipoproteiny E genetika MeSH
- aterosklerotický plát genetika patologie terapie MeSH
- ateroskleróza genetika patologie terapie MeSH
- buněčná diferenciace genetika MeSH
- cévy cytologie MeSH
- chondrogeneze genetika MeSH
- cytokiny metabolismus MeSH
- glykosaminoglykany metabolismus MeSH
- interleukin-6 metabolismus MeSH
- lidé MeSH
- mezenchymální kmenové buňky * MeSH
- myši MeSH
- osteogeneze genetika MeSH
- vaskulární kalcifikace genetika metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- ELISA metody přístrojové vybavení využití MeSH
- intracelulární signální peptidy a proteiny diagnostické užití izolace a purifikace krev MeSH
- klinické laboratorní techniky přístrojové vybavení využití MeSH
- lidé MeSH
- osteogeneze fyziologie genetika MeSH
- referenční standardy MeSH
- resorpce kosti genetika krev MeSH
- řízení kvality MeSH
- rozložení podle pohlaví MeSH
- statistika jako téma MeSH
- zdravotnické prostředky MeSH
- Check Tag
- lidé MeSH
Úvod: I když je vrozený defekt femuru (PFFD) dobře charakterizován morfologicky, změny na molekulární úrovni nejsou v literatuře popsány. Tento informační nedostatek společně s neznámou etiologií vady nás vedl k myšlence analýzy procesů angiogeneze a osteogeneze v tkáni pakloubu pacientů s PFFD v porovnání s fyziologickou kostí. Očekávali jsme rozdíly v genové expresi, zejména kvantitativní. Materiál a metodika: Během plánovaných ortopedických výkonů jsme odebrali kostní bloček, vložili do prezervačního média (RNA stabilization reagent), které brání degradaci RNA, a hluboce zmrazili. Poté jsme izolovali RNA a transkripční profil studovali biočipovou analýzou (SuperArray Bioscience Corporation). Celkem je tímto způsobem možno detekovat 113 genů osteogeneze a 113 genů angiogeneze. Od října roku 2005 do konce roku 2008 jsme vyšetřili vzorky od 7 pacientů s PFFD a 3 kontrolní vzorky. Některé analýzy jsme pro ověření výsledků opakovali, celkem jsme použili 13 čipů pro osteogenezi a 11 pro angiogenezi. Výsledky: Zaznamenali jsme rozdíly v kvantitě i typech exprimovaných genů. Některé geny byly exprimovány u PFFD více proti kontrolnímu vzorku (gen pro kalcitoninový receptor, kolagen XII, kolagen I alfa2, kolagen II, kolagen IX, FGFR2, fibronektin, integrin), jiné naopak vykazovaly sníženou expresi (gen pro annexin A5, kolagen XVIII alfa 1, kolagen I alfa1, katepsin K, FGFR1, FGFR3, IGF2, VEGFB). Závěr: Naše původní předpoklady se potvrdily rozdílnou genovou expresí. Prozatím získané výsledky nedovolují zobecnění, jsou pouze prvním krokem k dalším experimentům, kterými je třeba potvrdit avizované informace a propojit je s klinickými nálezy, jako je alternativní cévní zásobení postižené končetiny u některých pacientů.
Not only are teeth essential for mastication, but also missing teeth are considered a social handicap due to speech and aesthetic problems, with a resulting high impact on emotional well-being. Several treatment procedures are currently available for tooth replacement with mostly inert prosthetic materials and implants. Natural tooth substitution based on copying the developmental process of tooth formation is particularly challenging and creates a rapidly developing area of molecular dentistry. In any approach, functional interactions among the tooth, the surrounding bone, and the periodontium must be established. Therefore, recent research in craniofacial genetics searches for mechanisms responsible for correct cell and tissue interactions, not only within a specific structure, but also in the context of supporting structures. A tooth crown that is not functionally anchored to roots and bone is useless. This review aims to summarize the developmental and tissue homeostatic aspects of the tooth-bone interface, from the initial patterning toward tooth eruption and lifelong interactions between the tooth and its surrounding alveolar bone.
- MeSH
- homeoboxové geny MeSH
- lidé MeSH
- odontogeneze genetika MeSH
- osteoblasty fyziologie MeSH
- osteogeneze genetika MeSH
- periodontální vaz embryologie MeSH
- processus alveolaris embryologie MeSH
- prořezávání zubů MeSH
- signální transdukce MeSH
- zubní cement (tkáň) fyziologie MeSH
- zubní kořen embryologie MeSH
- zubní korunka (anatomie) embryologie MeSH
- zubní zárodek embryologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
