Biological Trace Element Research

, Volume 184, Issue 1, pp 136–147 | Cite as

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

  • Xingyu Zhu
  • Qianqian Gao
  • Genhua Zhao
  • Heng Wang
  • Ling Liu
  • Zhipeng Chen
  • Yijun Chen
  • Li Wu
  • Zisheng Xu
  • Weidong Li


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.


Raw pyritum Processed pyritum Bone healing Rat tibia defect TGF-β1/Smad signaling pathway 



Bone mineral density


Inductively coupled plasma-optical emission spectrometer


Alkaline phosphatase

Col I

Type I collagen




Transforming growth factor β


Traditional Chinese medicine


Reverse transcription polymerase chain reaction




Alpha modified eagle medium


Phosphate buffer solution


Fetal bovine serum


Glyceraldehyde-phosphate dehydrogenase


Tris-buffered saline Tween


Serum Ca


Serum P



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

Compliance with Ethical Standards

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.

Conflict of Interest

The authors declare that they have no conflicts of interest.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Xingyu Zhu
    • 1
  • Qianqian Gao
    • 1
  • Genhua Zhao
    • 1
  • Heng Wang
    • 1
  • Ling Liu
    • 1
  • Zhipeng Chen
    • 1
    • 2
  • Yijun Chen
    • 3
  • Li Wu
    • 1
  • Zisheng Xu
    • 4
  • Weidong Li
    • 1
    • 2
  1. 1.College of PharmacyNanjing University of Chinese MedicineNanjingPeople’s Republic of China
  2. 2.Engineering Center of State Ministry of Education for Standardization of Chinese Medicine ProcessingNanjing University of Chinese MedicineNanjingChina
  3. 3.Modern Analysis Center of Nanjing UniversityNanjingChina
  4. 4.Wuhu Pure Sunshine Natural Medicine Company LimitedWuhuPeople’s Republic of China

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