scholarly journals A Transgenic Rat for Noninvasive Assessment of Chondrogenesis in Vivo

Cartilage ◽  
2021 ◽  
pp. 194760352110572
Author(s):  
Elisabeth Ferreira ◽  
Landon B. Gatrell ◽  
Luke Childress ◽  
Hong Wu ◽  
Ryan M. Porter
Keyword(s):  
Bone Formation ◽  
Heterotopic Ossification ◽  
Ex Vivo ◽  
Firefly Luciferase ◽  
Bone Morphogenetic Protein 2 ◽  
Endochondral Bone Formation ◽  
Transgenic Rat ◽  
Regulatory Sequences ◽  
Endochondral Bone

Objective To support the preclinical evaluation of therapeutics that target chondrogenesis, our goal was to generate a rat strain that can noninvasively report endogenous chondrogenic activity. Design A transgene was constructed in which the dual expression of bioluminescent (firefly luciferase) and fluorescent (mCherry) reporters is controlled by regulatory sequences from rat Col2a1. Candidate lines were established on a Lewis background and characterized by serial bioluminescence imaging as well as ex vivo measurement of molecular reporter levels in several tissues. The sensitivity and specificity of the reporter strain were assessed in models of orthotopic and ectopic chondrogenesis. Results Substantial bioluminescence signal was detected from cartilaginous regions, including the appendicular synovial joints, spine, sternum, nose, and pinnae. Bioluminescent radiance was intense at 1 month of age, rapidly declined with continued development, yet remained detectable in 2-year-old animals. Explant imaging and immunohistochemistry confirmed that both molecular reporters were localized to cartilage. Implantation of wild-type bone marrow stromal cells into osteochondral defects made in both young adult and aged reporter rats led to a time-dependent elevation of intra-articular reporter activity concurrent with cartilaginous tissue repair. To stimulate ectopic, endochondral bone formation, bone morphogenetic protein 2 was overexpressed in the gastrocnemius muscle, which led to bioluminescent signal that closely preceded heterotopic ossification. Conclusions This strain can help develop strategies to stimulate cartilage repair and endochondral bone formation or to inhibit chondrogenesis associated with heterotopic ossification.

In Vivo Endochondral Bone Formation Using a Bone Morphogenetic Protein 2 Adenoviral Vector

1999 ◽  
Vol 10 (13) ◽  
pp. 2245-2253 ◽  
Author(s):  
Tord D. Alden ◽  
Debra D. Pittman ◽  
Gerald R. Hankins ◽  
Elisa J. Beres ◽  
Johnathan A. Engh ◽  
...  
Keyword(s):  
Bone Formation ◽  
Bone Morphogenetic Protein ◽  
Adenoviral Vector ◽  
Bone Morphogenetic Protein 2 ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Morphogenetic Protein

Effects of in vitro chondrogenic priming time of bone-marrow-derived mesenchymal stromal cells on in vivo endochondral bone formation

2015 ◽  
Vol 13 ◽  
pp. 254-265 ◽  
Author(s):  
Wanxun Yang ◽  
Sanne K. Both ◽  
Gerjo J.V.M. van Osch ◽  
Yining Wang ◽  
John A. Jansen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Endochondral Bone Formation ◽  
Endochondral Bone

scholarly journals Effects of Eleutherococcus Extract Mixture on Endochondral Bone Formation in Rats

2019 ◽  
Vol 20 (5) ◽  
pp. 1253 ◽  
Author(s):  
Donghun Lee ◽  
Sung Lee ◽  
Namhoon Cho ◽  
Young-Sik Kim ◽  
Jungbin Song ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Factor ◽  
Bone Formation ◽  
Female Adolescent ◽  
Bone Morphogenetic Protein 2 ◽  
Endochondral Bone Formation ◽  
Insulin Like Growth Factor ◽  
Endochondral Bone ◽  
Fluorescent Band ◽  
Traditional Medicines

Eleutherococcus extract mixture (EEM) is an herbal mixture of dried stem of Eleutherococcus sessiliflorus and germinated barley, which has been highly effective, in previous screening and among the traditional medicines to tonify innate qi and acquired qi, respectively. In this study, we investigate the effects of EEM on endochondral bone formation. Female adolescent rats were given EEM, growth hormone or vehicle for 10 days. Tetracycline was intraperitoneally injected to light the fluorescent band 72 h before sacrifice to determine endochondral bone formation. In order to evaluate endocrine or paracrine/autocrine mechanisms, expressions of insulin-like growth factor 1 (IGF1), insulin-like growth factor binding protein 3 (IGFBP3), or bone morphogenetic protein 2 (BMP2) were evaluated after EEM administration in liver or growth plate (GP). EEM oral administration significantly increased endochondral bone formation and proliferative and hypertrophic zonal heights of tibial GP. EEM also upregulated hepatic IGF1 and IGFBP3 mRNA expressions, and IGF1 and BMP2 expressions in GP. Taken together, EEM increases endochondral bone formation through stimulating proliferation and hypertrophy with upregulation of hepatic IGF1 and IGFBP3 expressions. Considering immunohistochemical studies, the effect of EEM may be due to increased local IGF1 and BMP2 expression in GP, which may be considered growth hormone (GH)-dependent endocrine and autocrine/paracrine pathways.

The transcription factor PRDM5 is involved in endochondral bone formation by regulating collagen I in vivo

Bone ◽  
2011 ◽  
Vol 48 ◽  
pp. S110
Author(s):  
G.G. Galli ◽  
K.H. de Lichtenberg ◽  
M. Wuelling ◽  
R. Calogero ◽  
A.H. Lund
Keyword(s):  
Transcription Factor ◽  
Bone Formation ◽  
Collagen I ◽  
Endochondral Bone Formation ◽  
Endochondral Bone

scholarly journals In Vitro and in Vivo Endochondral Bone Formation Models Allow Identification of Anti-Angiogenic Compounds

2003 ◽  
Vol 163 (1) ◽  
pp. 157-163 ◽  
Author(s):  
Gabri van der Pluijm ◽  
Martine Deckers ◽  
Bianca Sijmons ◽  
Henny de Groot ◽  
John Bird ◽  
...  
Keyword(s):  
Bone Formation ◽  
Endochondral Bone Formation ◽  
Endochondral Bone

scholarly journals Recombinant Vgr-1/BMP-6-expressing tumors induce fibrosis and endochondral bone formation in vivo.

1994 ◽  
Vol 126 (6) ◽  
pp. 1595-1609 ◽  
Author(s):  
S E Gitelman ◽  
M S Kobrin ◽  
J Q Ye ◽  
A R Lopez ◽  
A Lee ◽  
...  
Keyword(s):  
Bone Formation ◽  
Tumor Cells ◽  
Cho Cells ◽  
Subcutaneous Tissue ◽  
Endochondral Bone Formation ◽  
Related Factor ◽  
Tgf Beta ◽  
Endochondral Bone ◽  
Hypertrophic Cartilage

Members of the TGF-beta superfamily appear to modulate mesenchymal differentiation, including the processes of cartilage and bone formation. Nothing is yet known about the function of the TGF-beta-related factor vgr-1, also called bone morphogenetic protein-6 (BMP-6), and only limited studies have been conducted on the most closely related factors BMP-5, osteogenic protein-1 (OP-1) or BMP-7, and OP-2. Because vgr-1 mRNA has been localized in hypertrophic cartilage, this factor may play a vital role in endochondral bone formation. We developed antibodies to vgr-1, and documented that vgr-1 protein was expressed in hypertrophic cartilage of mice. To further characterize the role of this protein in bone differentiation, we generated CHO cells that overexpressed recombinant murine vgr-1 protein. Western blot analysis documented that recombinant vgr-1 protein was secreted into the media and was proteolytically processed to yield the mature vgr-1 molecule. To assess the biological activity of recombinant vgr-1 in vivo, we introduced the vgr-1-expressing CHO cells directly into the subcutaneous tissue of athymic nude mice. CHO-vgr-1 cells produced localized tumors, and the continuous secretion of vgr-1 resulted in tumors with a strikingly different gross and histological appearance as compared to the parental CHO cells. The tumors of control CHO cells were hemorrhagic, necrotic, and friable, whereas the CHO-vgr-1 tumors were dense, firm, and fibrotic. In contrast with control CHO tumors, the nests of CHO-vgr-1 tumor cells were surrounded by extensive connective tissue, which contained large regions of cartilage and bone. Further analysis indicated that secretion of vgr-1 from the transfected CHO tumor cells induced the surrounding host mesenchymal cells to develop along the endochondral bone pathway. These findings suggest that endochondral bone formation.

scholarly journals Bone Marrow Mesenchymal Stem Cells Form Ectopic Woven Bone In Vivo Through Endochondral Bone Formation

2009 ◽  
Vol 33 (4) ◽  
pp. 301-308 ◽  
Author(s):  
Sophia Chia-Ning Chang ◽  
Ching-Lung Tai ◽  
Hui-Ying Chung ◽  
Tsung-Min Lin ◽  
Long-Bin Jeng
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Woven Bone ◽  
Marrow Mesenchymal Stem Cells

scholarly journals Selective Deficiency of the “Bone-related”Runx2-II Unexpectedly Preserves Osteoblast-mediated Skeletogenesis

2004 ◽  
Vol 279 (19) ◽  
pp. 20307-20313 ◽  
Author(s):  
Zhou-Sheng Xiao ◽  
Anita B. Hjelmeland ◽  
L. D. Quarles
Keyword(s):  
Bone Formation ◽  
Ex Vivo ◽  
Mesenchymal Cells ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Chondrocyte Maturation ◽  
Intramembranous Bone ◽  
Exon 1 ◽  
Metaphyseal Bone ◽  
Deficient Mice

Runx2 (runt-related transcription factor 2) is a master regulator of skeletogenesis. Distinct promoters in the Runx2 gene transcribe the “bone-related”Runx2-II and non-osseousRunx2-I isoforms that differ only in their respective N termini. Existing mutant mouse models with both isoforms deleted exhibit an arrest of osteoblast and chondrocyte maturation and the complete absence of mineralized bone, but they do not distinguish the separate functions of the two N-terminal isoforms. To elucidate the function of the bone-related isoform, we generated selectiveRunx2-II-deficient mice by the targeted deletion of the distal promoter and exon 1. HomozygousRunx2-II-deficient (Runx2-II-/-) mice unexpectedly formed axial, appendicular, and craniofacial bones derived from either intramembranous ossification or mesenchymal cells of the bone collar, but they failed to form the posterior cranium and other bones derived from endochondral ossification. HeterozygousRunx2-II-deficient mice had grossly normal skeletons, but were osteopenic. The commitment of mesenchymal cellsex vivoto the osteoblast lineage occurred inRunx2-II-/-mice, but osteoblastic gene expression was impaired. Chondrocyte maturation appeared normal, but the zone of hypertrophic chondrocytes was not transformed into metaphyseal bone, leading to widened growth plates inRunx2-II-/-mice. Compensatory increments inRunx2-I expression occurred inRunx2-II-/-mice but were not sufficient to normalize osteoblastic maturation or transcriptional activity. Our findings support distinct functions ofRunx2-II and -I in the control of skeletogenesis.Runx2-I is sufficient for early osteoblastogenesis and intramembranous bone formation, whereasRunx2-II is necessary for complete osteoblastic maturation and endochondral bone formation.

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