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Table 1.

Primer sequences for quantitative real-time PCR.

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Fig 1.

PBE promoted osteogenic differentiation of rBMSCs in vitro.

(a) The ALP staining was conducted after 3-day treatment of PBE. The mineralization potential of rBMSCs was tested by Alizarin Red S staining after 7 days of PBE treatment. (b) Alizarin red S concentrations were quantified by absorbance measurement at 570 nM. (c) The genes expression of osteogenesis-related markers was assessed by quantitative real-time PCR after treatment of PBE after osteogenic induction for 3 and 7 days. * p < 0.05, ** p < 0.01, compared to the α-MEM group; # p < 0.05, ## p < 0.01, compared to the OIM group.

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Fig 2.

Animal experimental protocol (a) and representative X-rays (b) of distraction regenerate at various time points were present.

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Fig 3.

PBE treatment improved the quality of new callus as shown by μCT analysis and mechanical test.

(a) 3D μCT images of the tibia distraction zone in the two groups at week 3 and 6. (b) The value of BV/TV at week 3 and 6. (c) Mechanical properties (including E-modulus, ultimate load, and energy to failure) of distraction regenerates. *p < 0.05, compared to the PBS group, n = 4 at week 3; and n = 6 at week 6.

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Fig 4.

PBE adminstration accelerated new callus consolidation as shown by histological analysis.

(a) Representative sections stained with Goldner Trichrome. (b) Von Kossa staining.

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Fig 5.

Dynamic histomorphometric measurements showed more quantitative bone formation in the PBE treatment group.

(a) Arrows pointed to the Calcein and Xylenol orange labeling in representative images of two groups at week 3 and 6. (b) Quantitative measurements of dynamic histomorphometric parameters of MS/BS, MAR, BFR/BS, BFR/BV, and BFR/TV. *p< 0.05, ** p < 0.01, compared to the PBS group.

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Fig 6.

Representative images of immunohistochemical results of Osx (a-e) and OCN (a’-e’) and quantitative analysis of the positive cells in the distraction regenerates.

** p< 0.01, compared to the PBS group.

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