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Comparison of Bone Marrow Mesenchymal Stem Cells from Limb and Jaw Bones

Lloyd, Brandon R

Abstract Details

2016, Master of Science, Ohio State University, Dentistry.
Objectives: Bone marrow mesenchymal stem cells (BM-MSCs) from limb bones have demonstrated promises in regenerating craniofacial bones; yet little is known about the potential of BM-MSCs from craniofacial bones. This study compared BM-MSCs isolated from limb and craniofacial bones in a commonly used preclinical large animal model, the pig. Methods: Bone marrow was aspirated from the tibia and mandible (symphysis) of 4-month-old pigs (n=4). Subsequently, BM-MSCs were isolated, expanded and confirmed by flow cytometry. To assess cell proliferation, cell doubling times were calculated from serial cell number counts over 2 weeks. Total mRNA was extracted from freshly isolated BM-MSCs and analyzed for gene expression using an Affymetrix GeneChip porcine genome array, followed by real-time RT-PCR for validation. Osteogenic capacity was assessed by quantifying alkaline phosphatase activity. Using temperature-responsive culturing plates, the abilities of BM-MSCs to form multi-layer cell sheets (for future in-vivo transplantation), along with cell viability and morphology, were evaluated by fluorescent labeling and histological staining. Results: BM-MSCs from both locations expressed MSC markers but not hematopoietic markers. Mandibular BM-MSCs proliferated significantly faster than tibial BM-MSCs (median cell doubling times: 2.25 vs. 2.80 days, Mann-Whitney U test, p<0.01). Without any osteogenic induction, mandibular BM-MSC alkaline phosphatase activities were 3.3-fold (factorial ANOVA, p<0.001) to those of tibial BM-MSCs. Microarray analysis on one hand confirmed that overall mandibular and tibial BM-MSC gene expressions are highly correlated with each other and genes related to osteogenesis and angiogenesis were strongly expressed, and on the other hand revealed that there were several dozen genes which were indeed differentially expressed between mandibular and tibial BM-MSCs. They include cranial neural crest-related genes nestin (1.23-fold) and BMP-4 (1.79-fold), which were higher in mandibular BM-MSCs (ANOVA, p<0.05), a trend also confirmed by real-time RT-PCR tests. Three-layer cell sheets were successfully fabricated using both sources of BM-MSCs, but mandibular BM-MSCs required lower initial seeding density and had fewer dead cells than tibial BM-MSCs (3.13% vs. 10.25%, t-test, p<0.05). Conclusions: These data indicate that mandibular BM-MSCs may possess a greater potential than limb bone BM-MSCs for craniofacial bone regeneration.
Zongyang Sun, DDS/PhD/MS/MSD (Advisor)
85 p.

Recommended Citations

Citations

  • Lloyd, B. R. (2016). Comparison of Bone Marrow Mesenchymal Stem Cells from Limb and Jaw Bones [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1458678153

    APA Style (7th edition)

  • Lloyd, Brandon. Comparison of Bone Marrow Mesenchymal Stem Cells from Limb and Jaw Bones. 2016. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1458678153.

    MLA Style (8th edition)

  • Lloyd, Brandon. "Comparison of Bone Marrow Mesenchymal Stem Cells from Limb and Jaw Bones." Master's thesis, Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1458678153

    Chicago Manual of Style (17th edition)