Transplantation of Embryonic Fibroblasts Treated with Platelet-Rich Plasma Induces Osteogenesis in SAMP8 Mice Monitored by Molecular Imaging

Abstract

The aim of this study was to develop a cell-based boneregeneration approach evaluated by molecular imaging and immunohistochemistry. Methods: Genetically modified NIH3T3 embryonic fibroblasts carrying enhanced green fluorescent protein (NIH3T3-G) were predifferentiated into osteoblastlike cells using platelet-rich plasma (PRP) medium, followed by intraosseous transplantation into ovariectomized senescenceaccelerated mouse prone substrain 8 (OVX-SAMP8 mice). Results: PRP-conditioned NIH3T3-G (PRP/NIH3T3-G) engraftment prevented the development of osteoporosis. Molecular imaging and immunohistochemistry demonstrated the migration of NIH3T3-G cells from the implantation site throughout the skeleton. In situ analyses revealed coexpression of osteopontin and green fluorescent protein in the newly formed bone tissue, demonstrating that the transplant restored the bone trabecular architecture and mineral density in treated OVX-SAMP8 mice. Interestingly, the life span of OVX-SAMP8 mice receiving PRP/ NIH3T3-G transplantation was significantly prolonged and similar to that of the congenic senescence-resistant strain of mice. Conclusion: This unique and yet simple approach could potentially be applied to the treatment of senile postmenopausal osteoporosis and perhaps inborn genetic syndromes associated with accelerated aging, such as Hutchinson-Gilford progeria syndrome, and for the prolongation of life expectancy in general.

Delayed animal aging through the recovery of stem cell senescence by platelet rich plasma

“…To ensure the most appropriate concentration and consistency of the purified PRP used in this study, transforming growth factor beta 1 (TGF-b1) was quantitatively analyzed using a Quantikine enzymelinked immunosorbent assay (ELISA) kit (#DB100, R&D Diagnostics, Wiesbaden, Germany) to calibrate the purified PRP concentration. Previously, results showed that a 750 pg/ml concentration of TGF-b1 in PRP was optimal for cell proliferation and differentiation [20]. Purified PRP was dissolved with alpha-minimum essential medium (alpha-MEM) media (containing 1% fetal bovine serum and 1% prostatespecific antigen) to prepare the PRP solution for use in this study.…”

“…Previously, we demonstrated that platelet rich plasma (PRP) extended life span, improved survival and decreased adipogenesis in SAMP8 mice [19,20]. In this study, we hypothesized that PRP was able to delay aging through examining the characteristics of cells including cell growth (cell proliferation and colony formation), cell differentiation (osteogenesis and adipogenesis) and cellular senescence (gene, SA-b-gal activity, and oxidative stress); as well as animal morphology including life span, weight change and phenotype examination such as morphology (skin glossiness, collagen density, cataracts and lordokyphosis of the spine), bone formation and behavior (passivity and reactivity).…”

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

“…14). as the cells are capable of differentiating into various cell types such as neural cells, oesteocytes, and adipocytes, including insulin-producing beta cells [1,[19][20][21][24][25][26].…”

“…However, this group neither examined nor commented on the multipotent nature of these cells. Later, in 2008 and 2009, Deng and colleagues published two reports demonstrating differentiation of untransformed NIH3T3 cells into oesteocytes and neuronal cell types in bone and spinal cord injury mouse models [1,23,24]. Two other groups also reported that NIH3T3 can give rise to neural cells and oesteoblasts, respectively [21,25].…”

Tooth Loss Early in Life Accelerates Age-Related Bone Deterioration in Mice

“…SAMP8 mice show an accelerated senescence and ageassociated pathologies such as deficits in learning and memory, brain atrophy (Takeda et al 1994). SAMP8 mice were also used to mimic senile osteoporosis (Derave et al 2005;Lo et al 2009;Wauquier et al 2012;Furuzawa et al 2014). In this light, SAMP8 mice correctly mimic aging features and pathophysiologic environment regarding senile osteoporotic establishment in humans.…”