Histone deacetylase 4 deletion results in abnormal chondrocyte hypertrophy and premature ossification from collagen type 2α1‑expressing cells

Du,Guoqing; Xiang,Chuan; Sang,Xiaowen; Wang,Xiang; Shi,Ying; Wang,Nan; Wang,Shaowei; Li,Pengcui; Wei,Xiaochun; Zhang,Min; Gao,Lilan; Zhan,Hongsheng; Wei,Lei

doi:10.3892/mmr.2020.11465

Histone deacetylase 4 deletion results in abnormal chondrocyte hypertrophy and premature ossification from collagen type 2α1‑expressing cells

Authors:
- Guoqing Du
- Chuan Xiang
- Xiaowen Sang
- Xiang Wang
- Ying Shi
- Nan Wang
- Shaowei Wang
- Pengcui Li
- Xiaochun Wei
- Min Zhang
- Lilan Gao
- Hongsheng Zhan
- Lei Wei
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Affiliations: Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine (TCM), Institute of Traumatology and Orthopedics, Shanghai Academy of TCM, Shanghai 201203, P.R. China, Department of Orthopedics, The Second Hospital Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Tianjin Key Laboratory for Control Theory and Applications in Complicated Industry Systems, School of Mechanical Engineering, Tianjin University of Technology, Tianjin 300384, P.R. China, Department of Orthopedics, The Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, RI 02903, USA
Published online on: August 27, 2020 https://doi.org/10.3892/mmr.2020.11465
Pages: 4031-4040

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Abstract

Histone deacetylase 4 (HDAC4) plays a vital role in chondrocyte hypertrophy and bone formation. To investigate the function of HDAC4 in postnatal skeletal development, the present study developed lineage‑specific HDAC4‑knockout mice [collagen type 2α1 (Col2α1)‑Cre, HDAC4d/d mice] by crossing transgenic mice expressing Cre recombinase. Thus, a specific ablation of HDAC4 was performed in Col2α1‑expressing mice cells. The knee joints of HDAC4fl/fl and Col2α1‑Cre, HDAC4d/d mice were analyzed at postnatal day (P)2‑P21 using an in vivo bromodeoxyuridine (BrdU) assay, and Safranin O, Von Kossa and whole‑body staining were used to evaluate the developmental growth plate, hypertrophic differentiation, mineralization and skeletal mineralization patterns. The trabecular bone was analyzed using microcomputed tomography. The expressions of BrdU, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase (MMP)‑13, runt‑related transcription factor (Runx)‑2, osteoprotegerin (OPG), CD34, type X collagen (ColX), osteocalcin and Wnt5a were determined using immunohistochemistry, in situ hybridization (ISH) and reverse transcription‑quantitative (RT‑q)PCR. The results demonstrated that HDAC4‑null mice (HDAC4d/d mice) were severely runted; these mice had a shortened hypertrophic zone (histopathological evaluation), accelerated vascular invasion and articular mineralization (Von Kossa staining), elevated expressions of MMP‑13, Runx2, OPG and CD34 (RT‑qPCR and immunohistochemistry), downregulated expression of the proliferative marker BrdU and PCNA (immunohistochemistry), increased expression of ColX and decreased expression of Wnt5a (ISH). In conclusion, chondrocyte‑derived HDAC4 was responsible for regulating chondrocyte proliferation and differentiation as well as endochondral bone formation.

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