Advanced search options

Advanced Search Options 🞨

Browse by author name (“Author name starts with…”).

Find ETDs with:

in
/  
in
/  
in
/  
in

Written in Published in Earliest date Latest date

Sorted by

Results per page:

You searched for +publisher:"Temple University" +contributor:("Monroy, Maria A.;"). One record found.

Search Limiters

Last 2 Years | English Only

No search limiters apply to these results.

▼ Search Limiters


Temple University

1. Mundy, Christina Maria. The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function.

Degree: PhD, 2014, Temple University

Cell Biology

Connective tissue growth factor (CTGF/CCN2) and bone morphogenetic protein (BMP)-2 are both produced and secreted by osteoblasts. Both proteins have been shown to have independent effects in regulating osteoblast proliferation, maturation and mineralization. However, how these two proteins interact during osteoblast differentiation remains unknown. In Chapters 2 and 3, we utilized two cell culture model systems, osteoblasts derived from CTGF knockout (KO) mice and osteoblasts infected with an adenovirus, which over-expresses CTGF (Ad-CTGF), to investigate the effects of CTGF and BMP-2 on osteoblast development and function in vitro. To observe differences in osteoblast maturation and mineralization, we performed alkaline phosphatase (ALP) staining and activity and alizarin red staining, respectively. Contrary to a previously published report, osteoblast maturation and mineralization were similar in osteogenic cultures derived from KO and wild type (WT) calvaria in the absence of BMP-2 stimulation. Interestingly, in KO and WT osteoblast cultures stimulated with BMP-2, the KO osteoblast cultures exhibited increased alkaline phosphatase staining and activity and had larger, fused nodules stained with alizarin red than WT osteoblast cultures. This increase in osteoblast differentiation was accompanied by increased protein levels of phosphorylated Smad 1/5/8 and mRNA expression levels of bone morphogenetic protein receptor Ib. These data confirm enhanced osteoblast maturation and mineralization in BMP-2 induced KO osteoblast cultures. We also examined osteoblast differentiation in cultures that were infected with Ad-CTGF and in control cultures. Continuous over-expression of CTGF resulted in decreased ALP staining and activity, alizarin red staining, and mRNA expression of osteoblast markers in both unstimulated and BMP-2 stimulated cultures. Impaired osteoblast differentiation in cultures over-expressing CTGF was accompanied by decreased protein levels of phosphorylated Smad 1/5/8. In addition to the functional assays that we performed on WT and KO osteoblast cultures, we performed ChIP assays to investigate differences in binding occupancy of transcription factors on the Runx2 and Osteocalcin promoters in BMP-2 induced WT and KO osteoblast cultures. We demonstrate that in BMP-2 induced WT and KO osteoblast cultures, there was greater Smad 1 and JunB occupancy on the Runx2 promoter and Runx2 occupancy on the Osteocalcin promoter in BMP-2 induced KO osteoblast cultures compared to WT cultures. Collectively, the data demonstrate that CTGF acts to negatively regulate BMP-2 induced signaling and osteoblast differentiation. In Chapter 4, we synthesized an active His-tagged BMP-2 recombinant protein to track surface binding of BMP-2 in CTGF WT and KO osteoblasts. We amplified mature BMP-2 in genomic DNA, which was inserted correctly into a pET-28b(+) vector. We ran a SDS-PAGE gel and stained with Coomassie blue to show that we successfully induced BMP-2 in bacteria cells, extracted the protein using urea, and…

Advisors/Committee Members: Popoff, Steven N.;, Abood, Mary Ellen, Denny, Michael F., Goldfinger, Lawrence, Litvin, Judith, Monroy, Maria A.;.

Subjects/Keywords: Cellular biology;

Record DetailsSimilar RecordsGoogle PlusoneFacebookTwitterCiteULikeMendeleyreddit

APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Mundy, C. M. (2014). The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function. (Doctoral Dissertation). Temple University. Retrieved from http://digital.library.temple.edu/u?/p245801coll10,269581

Chicago Manual of Style (16th Edition):

Mundy, Christina Maria. “The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function.” 2014. Doctoral Dissertation, Temple University. Accessed October 20, 2020. http://digital.library.temple.edu/u?/p245801coll10,269581.

MLA Handbook (7th Edition):

Mundy, Christina Maria. “The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function.” 2014. Web. 20 Oct 2020.

Vancouver:

Mundy CM. The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function. [Internet] [Doctoral dissertation]. Temple University; 2014. [cited 2020 Oct 20]. Available from: http://digital.library.temple.edu/u?/p245801coll10,269581.

Council of Science Editors:

Mundy CM. The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function. [Doctoral Dissertation]. Temple University; 2014. Available from: http://digital.library.temple.edu/u?/p245801coll10,269581

.