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Comparison Study of Bone Defect Healing Effect of Raw and Processed Pyritum in Rats

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Abstract

To evaluate and compare the effect of raw and processed pyritum on tibial defect healing, 32 male Sprague Dawley rats were randomly divided into four groups. After tibial defect, animals were produced and grouped: sham and control group were orally administrated with distilled water (1 mL/100 g), while treatment groups were given aqueous extracts of raw and processed pyritum (1.5 g/kg) for successive 42 days. Radiographic examination showed that bone defect healing effect of the treatment groups was obviously superior compared to that of the control group. Bone mineral density of whole tibia was increased significantly after treating with pyritum. Inductively coupled plasma-optical emission spectrometry showed that the contents of Ca, P, and Mg in callus significantly increased in the treatment groups comparing with the control. Moreover, serological analysis showed that the concentration of serum phosphorus of the treatment groups significantly increased compared with that of the control group. By in vitro study, we have evaluated the effects of drug-containing serum of raw and processed pyritum on osteoblasts. It was manifested that both the drug-containing sera of raw and processed pyritum significantly increased the mRNA levels of alkaline phosphatase and collagen type I. Protein levels of phosphorylated Smad2/3 also increased. The mRNA levels of osteocalcin and transforming growth factor β (TGF-β) type I and II receptors, as well as the protein levels of TGF-β1 in the processed groups, were higher than those in the control. In summary, both raw and processed pyritum-containing sera exhibited positive effects on osteoblasts, which maybe via the TGF-β1/Smad signaling pathway. Notably, the tibia defect healing effect of pyritum was significantly enhanced after processing.

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Abbreviations

BMD:

Bone mineral density

ICP-OES:

Inductively coupled plasma-optical emission spectrometer

ALP:

Alkaline phosphatase

Col I:

Type I collagen

OC:

Osteocalcin

TGF-β:

Transforming growth factor β

TCM:

Traditional Chinese medicine

RT-PCR:

Reverse transcription polymerase chain reaction

PTFE:

Polytetrafluoroethylene

α-MEM:

Alpha modified eagle medium

PBS:

Phosphate buffer solution

FBS:

Fetal bovine serum

GAPDH:

Glyceraldehyde-phosphate dehydrogenase

TBST:

Tris-buffered saline Tween

S-Ca:

Serum Ca

S-P:

Serum P

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (81373970) and Jiangsu Qinglan Project (2014).

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Author notes

  1. Xingyu Zhu and Qianqian Gao contributed equally to this work.

Authors and Affiliations

  1. College of Pharmacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Nanjing, 210023, Jiangsu, People’s Republic of China

    Xingyu Zhu, Qianqian Gao, Genhua Zhao, Heng Wang, Ling Liu, Zhipeng Chen, Li Wu & Weidong Li

  2. Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing, China

    Zhipeng Chen & Weidong Li

  3. Modern Analysis Center of Nanjing University, Nanjing, China

    Yijun Chen

  4. Wuhu Pure Sunshine Natural Medicine Company Limited, Wuhu, Anhui, People’s Republic of China

    Zisheng Xu

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  1. Xingyu Zhu
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  3. Genhua Zhao
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Corresponding authors

Correspondence to Zisheng Xu or Weidong Li.

Ethics declarations

The study was approved by the Nanjing University of Chinese Medicine Committee on Laboratory Animal Care, and all animals received humane care according to the National Institutes of Health (USA) guidelines. All possible efforts were made to minimize the animals’ suffering and to reduce the number of animals used.

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The authors declare that they have no conflicts of interest.

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Zhu, X., Gao, Q., Zhao, G. et al. Comparison Study of Bone Defect Healing Effect of Raw and Processed Pyritum in Rats. Biol Trace Elem Res 184, 136–147 (2018). https://doi.org/10.1007/s12011-017-1166-0

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  • DOI: https://doi.org/10.1007/s12011-017-1166-0

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