Pamidronate Disodium Leads to Bone Necrosis via Suppression of Wnt/β-Catenin Signaling in Human Bone Marrow Mesenchymal Stem Cells In Vitro

  • J Oral Maxillofac Surg. 2017 Oct;75(10):2135-2143. doi: 10.1016/j.joms.2017.03.016.
Yan Xu  1 ,  Jin Sun  2 ,  Xuewen Yang  3 ,  Yuehai Yu  4 ,  Huijing Mai  5 ,  Zubing Li  6
Affiliations
  • 1. Physician, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and the Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China; Department of Oral and Maxillofacial Trauma and Plastic Aesthetic Surgery, School & Hospital of Stomatology, Wuhan University, Wuhan, China; Department of Stomatology, Shenzhen Luohu People's Hospital, Guangdong, China.
  • 2. Resident, Department of Stomatology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Guangdong; Department of Stomatology, The Affiliated Shenzhen Maternity and Child Healthcare Hospital of the South Medical University, Guangdong, China.
  • 3. Professor, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and the Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China; Department of Oral and Maxillofacial Trauma and Plastic Aesthetic Surgery, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
  • 4. Director, Department of Stomatology, Shenzhen Luohu People's Hospital, Guangdong, China.
  • 5. Physician, Department of Stomatology, Shenzhen Luohu People's Hospital, Guangdong, China.
  • 6. Professor, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and the Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China; Department of Oral and Maxillofacial Trauma and Plastic Aesthetic Surgery, School & Hospital of Stomatology, Wuhan University, Wuhan, China. Electronic address: [email protected].
PMID: 28412267 DOI: 10.1016/j.joms.2017.03.016
Abstract

Purpose: Pamidronate disodium-associated bone necrosis is poorly understood at the cellular and molecular levels. This study proposes a pathway leading to the pamidronate disodium-mediated inhibition of osteogenic differentiation of human bone marrow mesenchymal stem cells (BMMSCs) derived from the mandible in vitro.

Materials and methods: Primary human BMMSCs were isolated from the mandible and marrow tissue. A proliferation assay was performed to determine the experimental concentration of pamidronate disodium. Alkaline Phosphatase (ALP) activity, ALP staining, and Alizarin red S (ARS) staining were assessed after treatment with pamidronate disodium (0, 0.1, 0.5, 1, 5, 10 μg/mL). Quantitative real-time polymerase chain reaction and western blotting specific for Wnt and β-catenin signaling genes or proteins were performed after treatment with pamidronate disodium 0.5 μg/mL. Wnt3a was used to observe the osteogenic differentiation of BMMSCs during treatment with pamidronate disodium 0.5 μg/mL.

Results: As expected, pamidronate disodium 1, 5, and 10 μg/ml were unfavorable for BMMSC growth (P < .05), whereas 0.1 and 0.5 μg/mL did not affect BMMSC growth (P ≥ .05). BMMSCs treated with pamidronate disodium 0.5 μg/mL had lower ALP activity, ALP staining, and ARS staining (P < .05), and BMMSCs treated with low concentrations (<0.5 μg/mL) of pamidronate disodium had the same levels of ALP activity, ALP staining, and ARS staining as the control (0 μg/mL). Pamidronate disodium 0.5 μg/mL decreased the expression of genes and proteins involved in Wnt and β-catenin signaling. BMMSCs with Wnt3a and pamidronate disodium 0.5 μg/mL had higher ALP activity, ALP staining, and ARS staining (P < .05).

Conclusions: Pamidronate disodium inhibited Wnt and β-catenin signaling, which controls osteogenic differentiation in BMMSCs. Wnt3a, a Wnt and β-catenin signaling activator, reversed the negative effects caused by pamidronate disodium to salvage the osteogenic defect in BMMSCs.

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