scholarly journals Runx2 is required for hypertrophic chondrocyte mediated degradation of cartilage matrix during endochondral ossification

2021 ◽  
pp. 100088
Author(s):  
Harunur Rashid ◽  
Haiyan Chen ◽  
Amjad Javed
Keyword(s):  
Endochondral Ossification ◽  
Cartilage Matrix ◽  
Hypertrophic Chondrocyte

scholarly journals Clumps of Mesenchymal Stem Cells/Extracellular Matrix Complexes Generated with Xeno-Free Chondro-Inductive Medium induce Bone Regeneration via Endochondral Ossification

Biomedicines ◽  
2021 ◽  
Vol 9 (10) ◽  
pp. 1408
Author(s):  
Susumu Horikoshi ◽  
Mikihito Kajiya ◽  
Souta Motoike ◽  
Mai Yoshino ◽  
Shin Morimoto ◽  
...  
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Bone Regeneration ◽  
Endochondral Ossification ◽  
Healing Process ◽  
Early Period ◽  
Cartilage Matrix

Three-dimensional clumps of mesenchymal stem cells (MSCs)/extracellular matrix (ECM) complexes (C-MSCs) can be transplanted into tissue defect site with no artificial scaffold. Importantly, most bone formation in the developing process or fracture healing proceeds via endochondral ossification. Accordingly, this present study investigated whether C-MSCs generated with chondro-inductive medium (CIM) can induce successful bone regeneration and assessed its healing process. Human bone marrow-derived MSCs were cultured with xeno-free/serum-free (XF) growth medium. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and then torn off. The sheet was rolled to make a round clump of cells. The cell clumps, i.e., C-MSCs, were maintained in XF-CIM. C-MSCs generated with XF-CIM showed enlarged round cells, cartilage matrix, and hypertrophic chondrocytes genes elevation in vitro. Transplantation of C-MSCs generated with XF-CIM induced successful bone regeneration in the SCID mouse calvaria defect model. Immunofluorescence staining for human-specific vimentin demonstrated that donor human and host mouse cells cooperatively contributed the bone formation. Besides, the replacement of the cartilage matrix into bone was observed in the early period. These findings suggested that cartilaginous C-MSCs generated with XF-CIM can induce bone regeneration via endochondral ossification.

Defective bone formation in Krox-20 mutant mice

Development ◽  
1996 ◽  
Vol 122 (1) ◽  
pp. 113-120 ◽  
Author(s):  
G. Levi ◽  
P. Topilko ◽  
S. Schneider-Maunoury ◽  
M. Lasagna ◽  
S. Mantero ◽  
...  
Keyword(s):  
Bone Formation ◽  
Endochondral Ossification ◽  
Axial Skeleton ◽  
Lacz Gene ◽  
E Coli ◽  
Hypertrophic Chondrocyte ◽  
Central Regions ◽  
Vertebrate Skeleton ◽  
Defective Bone ◽  
Skeleton Formation

Endochondral ossification is the prevalent mode of vertebrate skeleton formation; it starts during embryogenesis when cartilage models of long bones develop central regions of hypertrophy which are replaced by bony trabeculae and bone marrow. Although several transcription factors have been implicated in pattern formation in the limbs and axial skeleton, little is known about the transcriptional regulations involved in bone formation. We have created a null allele in the mouse Krox-20 gene, which encodes a zinc finger transcription factor, by in frame insertion of the E. coli lacZ gene and shown that hindbrain segmentation and peripheral nerve myelination are affected in Krox-20−/− embryos. We report here that Krox-20 is also activated in a subpopulation of growth plate hypertrophic chondrocytes and in differentiating osteoblasts and that its disruption severely affects endochondral ossification. Krox-20−/− mice develop skeletal abnormalities including a reduced length and thickness of newly formed bones, a drastic reduction of calcified trabeculae and severe porosity. The periosteal component to bone formation and calcification does not appear to be affected in the homozygous mutant suggesting that the major role for Krox-20 is to be found in the control of the hypertrophic chondrocyte-osteoblast interactions leading to endosteal bone formation.

scholarly journals Effect of Diet on Longitudinal Bone Growth and Osteochondrosis in Swine

1987 ◽  
Vol 24 (2) ◽  
pp. 109-117 ◽  
Author(s):  
J. C. Woodard ◽  
H. N. Becker ◽  
P. W. Poulos
Keyword(s):  
Articular Cartilage ◽  
Growth Plate ◽  
Radial Growth ◽  
Endochondral Ossification ◽  
Bone Growth ◽  
Cartilage Matrix ◽  
Protein Diet ◽  
Cell Production ◽  
Proliferative Zone ◽  
Daily Rate

Weanling gilts were fed either a 12% or 16% protein diet for 10 weeks. Animals fed the 12% protein diet had reduced body weights and reduced longitudinal bone growth as measured in the distal radial growth plate. There was no difference in the growth plate widths between the two animal groups, but there was a significant reduction in the daily rate of cell production in the proliferative zone of animals fed the 12% protein diet. No effect of diet on the rate of expansion of the epiphysis at the articular-epiphyseal junction of the distal femur or humerus could be detected. All animals in both groups had morphologic cartilage lesions consistent with early changes associated with osteochondrosis (OCD), and there was no difference in the lesion morphology between the dietary groups. Areas of disorderly endochondral ossification in the radial growth plate were associated with perpendicular growth cartilage infractions. Growth plate lesions were characterized by increased widths of the maturing cartilage zone without increased width of the proliferative zone or an increase in the daily rate of cell production. Focal growth plate lesions developed because of a transitory inhibition of cartilage mineralization and resorption. Disorderly foci of endochondral ossification beneath articular cartilage were characterized by an area of chondrocyte necrosis which prevented normal cartilage matrix mineralization. Lamellae of cartilage necrosis were also present within the reserve zone of the articular cartilage. These were associated with abnormalities of the cartilage canal vessels, and chondrocyte necrosis was considered to precede degenerative changes in articular cartilage matrix. Since similar changes were rarely seen in the distal metacarpal (tarsal) bones of the dew claws, endogenous trauma was considered an important factor in lesion pathogenesis. The possibility that a basic cartilage matrix abnormality might represent the primary cause of OCD is discussed.

scholarly journals Liquid-phase ASEM imaging of cellular and structural details in cartilage and bone formed during endochondral ossification: Keap1-deficient osteomalacia

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Eiko Sakai ◽  
Mari Sato ◽  
Nassirhadjy Memtily ◽  
Takayuki Tsukuba ◽  
Chikara Sato
Keyword(s):  
Cortical Bone ◽  
Endochondral Ossification ◽  
Soft Tissues ◽  
Phosphotungstic Acid ◽  
Developmental Stages ◽  
Bone Development ◽  
Radical Scavenger ◽  
Hypertrophic Chondrocyte ◽  
Structural Details ◽  
Bone Mineralisation

AbstractChondrogenesis and angiogenesis drive endochondral ossification. Using the atmospheric scanning electron microscopy (ASEM) without decalcification and dehydration, we directly imaged angiogenesis-driven ossification at different developmental stages shortly after aldehyde fixation, using aqueous radical scavenger glucose solution to preserve water-rich structures. An embryonic day 15.5 mouse femur was fixed and stained with phosphotungstic acid (PTA), and blood vessel penetration into the hypertrophic chondrocyte zone was visualised. We observed a novel envelope between the perichondrium and proliferating chondrocytes, which was lined with spindle-shaped cells that could be borderline chondrocytes. At postnatal day (P)1, trabecular and cortical bone mineralisation was imaged without staining. Additional PTA staining visualised surrounding soft tissues; filamentous connections between osteoblast-like cells and osteocytes in cortical bone were interpreted as the osteocytic lacunar-canalicular system. By P10, resorption pits had formed on the tibial trabecular bone surface. The applicability of ASEM for pathological analysis was addressed using knockout mice of Keap1, an oxidative-stress sensor. In Keap1−/− femurs, we observed impaired calcification and angiogenesis of epiphyseal cartilage, suggesting impaired bone development. Overall, the quick ASEM method we developed revealed mineralisation and new structures in wet bone tissue at EM resolution and can be used to study mineralisation-associated phenomena of any hydrated tissue.

scholarly journals Mutations in fam20b and xylt1 Reveal That Cartilage Matrix Controls Timing of Endochondral Ossification by Inhibiting Chondrocyte Maturation

PLoS Genetics ◽  
2011 ◽  
Vol 7 (8) ◽  
pp. e1002246 ◽  
Author(s):  
B. Frank Eames ◽  
Yi-Lin Yan ◽  
Mary E. Swartz ◽  
Daniel S. Levic ◽  
Ela W. Knapik ◽  
...  
Keyword(s):  
Endochondral Ossification ◽  
Cartilage Matrix ◽  
Chondrocyte Maturation

The Effects of Fluvastatin on Indian Hedgehog Pathway in Endochondral Ossification

Cartilage ◽  
2019 ◽  
pp. 194760351986231
Author(s):  
Munetada Ishikawa ◽  
Takenobu Ishii ◽  
Taiki Morikawa ◽  
Yuki Iijima ◽  
Kenji Sueishi
Keyword(s):  
Cell Proliferation ◽  
Endochondral Ossification ◽  
Low Density Lipoprotein ◽  
Growth Factor Receptor ◽  
Density Lipoprotein ◽  
Indian Hedgehog ◽  
Related Protein ◽  
Hypertrophic Chondrocyte ◽  
Parathyroid Hormone Related Protein ◽  
Induced Pluripotent

Statins have demonstrated to be effective for treating chondrodysplasia and its effects were believed to be associated with the fibroblast growth factor receptor 3 (FGFR3). Statins promoted the degradation of FGFR3 in studies using disease-specific induced pluripotent stem cells and model mice, however, recent studies using normal chondrocytes reported that statins did not degrade FGFR3. In order to further investigate the effects of statins in endochondral ossification, this study examined the influence of statins on Indian hedgehog (Ihh), another important component of endochondral ossification, and its related pathways. The chondrocyte cell line ATDC5 was used to investigate changes in cell proliferation, mRNA, and protein expression levels. In addition, an organ culture of a mouse metatarsal bone was performed followed by hematoxylin-eosin staining and fluorescent immunostaining. Results indicated that expression level of Ihh increased with the addition of statins, which activated the Ihh pathway and altered the localization of Ihh. Changes in cholesterol modification may have affected Ihh diffusibility; however, further experiments are necessary. A reactive increase in parathyroid hormone–related protein (PTHrP) was observed in addition to changes in the Wnt pathway through secreted-related protein 2/3 and low-density lipoprotein 5/6. This led to the promotion of cell proliferation, increase of the hypertrophic chondrocyte layer, inhibition of apoptosis, and decrease in mineralization. This study demonstrated that statins had an influence on Ihh, and that the hyperfunction of Ihh may prevent premature cell death caused by FGFR3-related chondrodysplasia through an indirect increase in the expression of PTHrP.

Effects of EDTA, Hyaluronidase and Collagenase on Epiphyseal Cartilage Matrix

1968 ◽  
Vol 26 ◽  
pp. 56-57
Author(s):  
H. Clarke Anderson ◽  
Priscilla R. Coulter
Keyword(s):  
Light And Electron Microscopy ◽  
Matrix Vesicles ◽  
Cartilage Matrix ◽  
Collagen Fibrils ◽  
Epiphyseal Cartilage ◽  
Dense Matrix ◽  
Type Iv ◽  
Bovine Testicular Hyaluronidase ◽  
The Matrix ◽  
Testicular Hyaluronidase

Epiphyseal cartilage matrix contains fibrils and particles of at least 5 different types: 1. Banded collagen fibrils, present throughout the matrix, but not seen in the lacunae. 2. Non-periodic fine fibrils <100Å in diameter (Fig. 1), which are most notable in the lacunae, and may represent immature collagen. 3. Electron dense matrix granules (Fig. 1) which are often attached to fine fibrils and collagen fibrils, and probably contain protein-polysaccharide although the possibility of a mineral content has not been excluded. 4. Matrix vesicles (Fig. 2) which show a selective distribution throughout the epiphysis, and may play a role in calcification. 5. Needle-like apatite crystals (Fig. 2).Blocks of formalin-fixed epiphysis from weanling mice were digested with the following agents in 0.1M phosphate buffer: a) 5% ethylenediaminetetraacetate (EDTA) at pH 8.3, b) 0.015% bovine testicular hyaluronidase (Sigma, type IV, 750 units/mg) at pH 5.5, and c) 0.1% collagenase (Worthington, chromatograhically pure, 200 units/mg) at pH 7.4. All digestions were carried out at 37°C overnight. Following digestion tissues were examined by light and electron microscopy to determine changes in the various fibrils and particles of the matrix.

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