Transcriptional analysis of human cranial compartments with different embryonic origins
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE62816 | GEO | 2015/08/03
SECONDARY ACCESSION(S): PRJNA265217
REPOSITORIES: GEO
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Project description:Previous investigations suggest that the different embryonic origins of the calvarial tissues (neural crest or mesoderm) may account for the different molecular mechanisms underlying sutural development. The aim of this study was to evaluate the differences in the gene expression of human cranial tissues and assess the presence of an expression signature reflecting their embryonic origins. Using microarray technology, we investigated global gene expression of cells from the frontal and parietal bones and the metopic and sagittal intrasutural mesenchyme (ISM) of four human fetal calvaria. Tissues samples were obtained from fetal crania of 4 normal human fetuses (two females ages 94 and 103 days and two males ages 97 and 98 days). All dural and extracranial soft tissues were removed from the parietal and frontal bones. 2-3 mm tissue compartments were excised from frontal and parietal bones and metopic and sagittal ISM. To avoid contamination with osteoblasts, ISM tissue was dissected from the central portion of each suture. Similarly, bone was harvested at least 3mm from the margin of the suture to avoid contamination with ISM. Media was changed every 3-4 days. After reaching 75-80% confluence, the cells were trypsinized with TrypLEExpress (Life Technologies, Denmark) and passaged. During the fourth and final passage, 180,000 cells were plated in triplicate in 6-well plates and cultured for 5 days followed by RNA extraction.
2015-08-03 | E-GEOD-62816 | biostudies-arrayexpress
Project description:Purpose: This study investigates the postnatal impact of Twist1 haploinsufficiency on the osteoskeletal ability and regeneration on two calvarial bones arising from tissues of different embryonic origin: the neural crest-derived frontal and the mesoderm-derived parietal bones. Results: Twist1 haplonsufficiency selectively enhanced osteogenic and tissue regeneration ability of mesoderm-derived parietal bones. Twist1 haplonsufficiency triggers its selective activity on mesoderm-derived parietal bone through downregulation of the bone-derived hormone Fgf23. Conclusions: Twist1 haploinsufficiency preferentially triggers osteogenic induction of parietal bones which may be beneficial to optimize treatments for skeletal regeneration, reconstruction and repair of mesoderm-derived bone, as well as to alleviate skeletal abnormalities caused by Twist1 haploinsufficiency.
2018-09-15 | GSE119962 | GEO
Project description:Background: It has widely been observed that young children are capable of reossifying large calvarial defects, while adults lack this endogenous tissue-engineering capacity. The ability of juvenile animals to regenerate calvarial defects has been investigated in multiple animal models, including mice. In this study, the authors used cDNA microarrays to investigate the expression of osteogenesis-associated genes upstream and downstream of Runx2 in juvenile and adult mouse calvaria. Methods: Nonsuture-associated parietal bone discs were harvested from 6-day-old (n = 50) and 60-day-old (n = 35) male CD-1 mice. After separation of the underlying dura mater and overlying pericranium, the calvarial discs were snap-frozen and RNA was extracted from pooled samples of calvaria for microarray analysis. Genes analyzed included cytokines, receptors, and cell-surface and matrix proteins both upstream and downstream of Runx2. Results: Genes associated with the Runx2 pathway had notably higher levels in the juvenile versus adult calvaria. All genes except for osteocalcin were expressed at least twofold higher in the juvenile calvaria. This pattern was validated with quantitative real-time polymerase chain reaction. In addition, mRNA for potent osteoinductive growth factors was present at higher levels in the juvenile compared with the adult calvaria. Conclusions: These findings reflect a genomic environment of active osteoblast differentia-tion and ossification in the juvenile calvaria compared with the adult aquiescent calvarial tissue. These data suggest that a decreased osteogenic potential of adult calvarial osteoblasts may, in part, explain the inability of adult animals to heal calvarial defects.
2005-07-05 | E-SMDB-3396 | biostudies-arrayexpress
Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a Porphyromonas gingivalis infection using a murine calvarial model of inflammation and bone resorption. P. gingivalis strain 381 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChip® MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process. P. gingivalis was injected at 1.5 x 10E9 (N = 10) into the soft tissues overlying the calvaria of the mice for 3 days. A control group (N = 9) was injected with vehicle once daily for 3 days. Mice were euthanized 8 h after the last injection. The calvarial bones and overlying soft tissues from 5 mice in each group were excised, snap frozen in liquid nitrogen, and stored at –80°C until RNA isolation. Total RNA was isolated from the frozen calvarial tissue and calvarial bone from each mouse (P. gingivalis infected and control animals) with Trizol reagent (Invitrogen, CA). Equal amounts of RNA from samples were labeled and hybridized on a mouse GeneChip following the protocol described in the GeneChip Expression Analysis Technical manual (Affymetrix, Santa Clara, CA). After hybridization, the GeneChip arrays were stained and scanned in an Affymetrix GCS 3000 7G Scanner.
2010-07-15 | E-GEOD-17110 | biostudies-arrayexpress
Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a Treponema denticola infection using a murine calvarial model of inflammation and bone resorption. T. denticola ATCC 35404 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChip® MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process. T. denticola was injected at 1.5 x 109 (N = 10) into the soft tissues overlying the calvaria of the mice for 3 days. A control group (N = 9) was injected with vehicle once daily for 3 days. Mice were euthanized 8 h after the last injection. The calvarial bones and overlying soft tissues from 5 mice in each group were excised, snap frozen in liquid nitrogen, and stored at –80°C until RNA isolation. Total RNA was isolated from the frozen calvarial tissue and calvarial bone from each mouse (T. denticola infected and control animals) with Trizol reagent (Invitrogen, CA). Equal amounts of RNA from samples were labeled and hybridized on a mouse GeneChip following the protocol described in the GeneChip Expression Analysis Technical manual (Affymetrix, Santa Clara, CA). After hybridization, the GeneChip arrays were stained and scanned in an Affymetrix GCS 3000 7G Scanner.
2010-11-16 | E-GEOD-19855 | biostudies-arrayexpress
Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a polymicrobial periodontal pathogens Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia infection using a murine calvarial model of inflammation and bone resorption. P. gingivalis FDC 381, T. denticola ATCC 35404, and T. forsythia ATCC 43037 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChipM-BM-. MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process. P. gingivalis/T. denticola/T. forsythia was injected at 5 x 108 each (N = 10) into the soft tissues overlying the calvaria of the mice for 3 days. A control group (N = 9) was injected with vehicle once daily for 3 days. Mice were euthanized 8 h after the last injection. The calvarial bones and overlying soft tissues from 5 mice in each group were excised, snap frozen in liquid nitrogen, and stored at M-bM-^@M-^S80M-BM-0C until RNA isolation. Total RNA was isolated from the frozen calvarial tissue and calvarial bone from each mouse (Polymicrobial bacteria infected and control animals, N = 5 in each group) with Trizol reagent (Invitrogen, CA). Equal amounts of RNA from samples were labeled and hybridized on a mouse GeneChip following the protocol described in the GeneChip Expression Analysis Technical manual (Affymetrix, Santa Clara, CA). After hybridization, the GeneChip arrays were stained and scanned in an Affymetrix GCS 3000 7G Scanner.
2014-01-10 | E-GEOD-29670 | biostudies-arrayexpress
Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a Treponema denticola infection using a murine calvarial model of inflammation and bone resorption. T. denticola ATCC 35404 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChip® MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process.
2010-11-16 | GSE19855 | GEO
Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a Tannerella forsythia infection using a murine calvarial model of inflammation and bone resorption. T. forsythia ATCC 43037 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChip® MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process.
2011-02-07 | GSE20389 | GEO
Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a Porphyromonas gingivalis infection using a murine calvarial model of inflammation and bone resorption. P. gingivalis strain 381 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChip® MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process.
2010-07-15 | GSE17110 | GEO
Project description:In newborn humans, and up to approximately 2 y of age, calvarial bone defects can naturally regenerate. This remarkable regeneration potential is also found in newborn mice and is absent in adult mice. Since previous studies showed that the mouse calvarial sutures are reservoirs of calvarial skeletal stem cells (cSSCs), which are the cells responsible for calvarial bone regeneration, here we hypothesized that the regenerative potential of the newborn mouse calvaria is due to a significant amount of cSSCs present in the newborn expanding sutures. Thus, we tested whether such regenerative potential can be reverse engineered in adult mice by artificially inducing an increase of the cSSCs resident within the adult calvarial sutures.First, we analyzed the cellular composition of the calvarial sutures in newborn and in older mice, up to 14-mo-old mice, showing that the sutures of the younger mice are enriched in cSSCs. Then, we demonstrated that a controlled mechanical expansion of the functionally closed sagittal sutures of adult mice induces a significant increase of the cSSCs. Finally, we showed that if a calvarial critical size bone defect is created simultaneously to the mechanical expansion of the sagittal suture, it fully regenerates without the need for additional therapeutic aids. Using a genetic blockade system, we further demonstrate that this endogenous regeneration is mediated by the canonical Wnt signaling.This study shows that controlled mechanical forces can harness the cSSCs and induce calvarial bone regeneration. Similar harnessing strategies may be used to develop novel and more effective bone regeneration autotherapies.
2023-04-11 | GSE227468 | GEO