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Figure 1.

Otolaryngologic finding of the nasal cavity.

A: Schematic of the lateral nasal wall of the right nasal cavity. B: Endoscopic visualization of the right nasal cavity. *Inferior turbinate.

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Table 1.

Oligonucleotide primer sequences used for reverse transcriptase polymerase chain reaction (RT-PCR).

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Table 2.

Effects of donor age on the immunophenotypic characteristics of hTMSCs.

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Table 3.

Effects of donor age on the neural crest markers of hTMSCs.

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Figure 2.

Proliferation of hTMSCs according to age group.

Cellular proliferation was monitored over a period of 14 days. hTMSCs from all groups exhibited rapid proliferation from day 2 to 4; hTMSCs from group (II) expanded more rapidly than those from the other groups. However, there were no statistically significant differences in proliferation rate among the groups.

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Figure 3.

Effect of donor age on the osteogenic differentiation potential of hTMSCs.

Cells were cultured in osteogenic induction medium. Cells aggregated, formed nodules, and accumulated calcium deposits over a 2-week period. A–D: Alkaline phosphatase staining of hTMSCs cultured from each age group before osteogenic induction. E–H: Alkaline phosphatase staining of hTMSCs cultured from each age group after osteogenic induction. Alkaline phosphatase activity, which is indicative of osteoblastic differentiation, is shown as red staining. I–L: Alizarin red staining of hTMSCs cultured from each age group after osteogenic induction. (A, E, and I: group I; B, F, and J: group II; C, G, and K: group III; D, H, and L: group IV). Alizarin Red staining was used to detect precipitated calcium salt, which is a marker of differentiation. Scale bars: 100 µm. Visual assessment data demonstrated that hTMSCs cultured from donors of all age groups showed the alkaline phosphatase activity increase after osteogenic induction and similar levels of alkaline phosphatase and Alizarin Red staining.

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Figure 4.

RT-PCR analysis of osteoblast-specific genes expression during 2 weeks of culture in osteogenically differentiated hTMSCs.

Bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP), type I collagen (Col1), osteocalcin (OC), osterix (Osx), and osteopontin (OP) mRNA expression were analyzed. The experiment was performed in triplicate for each sample. Differences among the groups were evaluated by one-way analysis of variance (ANOVA; p<0.05). Except for OP, there were no significant differences in expression levels among the groups.

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