Temporal expression of wound healing–related genes inform wound age estimation in rats after a skeletal muscle contusion: a multivariate statistical model analysis

  • Qiu-xiang Du
  • Na Li
  • Li-hong Dang
  • Ta-na Dong
  • Han-lin Lu
  • Fu-xia Shi
  • Qian-qian Jin
  • Cao JieEmail author
  • Jun-hong SunEmail author
Original Article


Although many time-dependent parameters involved in wound healing have been exhaustively investigated, establishing an objective and reliable means for estimating wound age remains a challenge. In this study, 78 Sprague–Dawley rats were divided randomly into a control group and contusion groups at 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and 48 h post-injury (n = 6 per group). The expression of 35 wound healing–related genes was explored in contused skeletal muscle by real-time polymerase chain reaction. Differences between the groups were assessed by partial least squares discriminant analysis (PLS-DA). The results show that the samples were classified into three groups by wound age (4–12, 16–24, and 28–48 h). A Fisher discriminant analysis model of 14 selected genes was constructed, and 94.9% cross-validated grouped cases were correctly classified. A PLS regression analysis using 14 genes showed reasonable internal predictive validity, with a root mean squared error of cross-validation of approximately 8 h. To examine whether the prediction models were capable of analyzing new (ungrouped) cases, an external validation was carried out using the expression data from an additional 30 rats. Approximately 76.7% of ungrouped cases were correctly classified, which was a lower proportion than that for cross-validation. Similarly, the prediction results of the PLS model showed lower relatively external predictive validity (root mean squared error of prediction = 11 h) than internal predictive validity. Although the prediction results were less accurate than expected, the gene expression modeling and multivariate analyses showed great potential for estimating injury time. These multivariate methods may be valuable when devising future wound time estimation strategies.


FDA Multivariate statistical model analysis PLSR Real-time PCR Skeletal muscle contusion Wound age estimation 


Funding information

This project is financially supported by grants from the National Natural Science Foundation of China (grant numbers 81601646, 81571852).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. The principles of the Guide for the Care and Use of Laboratory Animals protocol, published by the Ministry of the People’s Republic of China, were followed.

Supplementary material

414_2018_1990_MOESM1_ESM.docx (28 kb)
ESM 1 (DOCX 28 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Forensic MedicineShanxi Medical UniversityTaiyuanPeople’s Republic of China

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