Jian Wu1, Ji-Feng Huang2, Xing-Xing Qin3, Feng Hu3, Zhao-Fei Chen3, Yong Zheng3, Yan-Xi Liu3, Xian-Hua Cai4. 1. Southern Medical University, Guangzhou 510515, China; Department of Orthopaedics, XianNing Central Hospital, The First Affiliated Hospital of Hubei University of Science and Technology, XianNing 437100, China. 2. Department of Orthopaedics, Wuhan General Hospital of Guangzhou Military Area Commands affiliated Southern Medical University, Wuhan, China. 3. Department of Orthopaedics, XianNing Central Hospital, The First Affiliated Hospital of Hubei University of Science and Technology, XianNing 437100, China. 4. Southern Medical University, Guangzhou 510515, China; Department of Orthopaedics, Wuhan General Hospital of Guangzhou Military Area Commands affiliated Southern Medical University, Wuhan, China. Electronic address: wgcaixh@163.com.
Abstract
OBJECTIVE: Platelet-rich plasma (PRP) has been reported to alleviate degenerative pathological damage to joint cartilage. This study aimed to investigate the effect of PRP on Wnt/β-catenin signaling in rabbit chondrocytes. METHODS: Using 3-month-old New Zealand white rabbits, PRP was prepared from venous blood, and chondrocytes were cultured from knee joint cartilage and identified by staining for type II collagen and proteoglycan. The effects of PRP on chondrocyte viability were measured. The chondrocytes were divided into 5 groups: control, IL-1β, PRP (100-fold dilution), Dkk-1 (100ng/mL) and Dkk-1+PRP. The IL-1β, PRP, Dkk-1 and Dkk-1+PRP groups were treated with interleukin (IL)-1β (50μL, 10μg/mL) for24h. Chondrocyte morphology was observed by electron microscopy. Levels of carboxy terminal peptide (CTX-II) and cartilage oligomeric matrix protein (COMP) in culture media were measured by ELISA. Wnt-1, β-catenin and GSK-3β mRNA and protein expression were determined by RT-PCR and western blot respectively. RESULTS: PRP enhanced chondrocyte proliferation. Chondrocytes in the IL-1β group showed ultrastructural abnormalities that were less pronounced in the PRP, Dkk-1 and Dkk-1+PRP groups. CTX-II and COMP concentrations were higher in the IL-1β group than in the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). The IL-1β group had higher mRNA and protein Wnt1 and β-catenin levels and lower GSK-3β levels than the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). CONCLUSION: PRP may protect chondrocytes activated by IL-1β via inhibiting Wnt/β-catenin signaling.
OBJECTIVE: Platelet-rich plasma (PRP) has been reported to alleviate degenerative pathological damage to joint cartilage. This study aimed to investigate the effect of PRP on Wnt/β-catenin signaling in rabbit chondrocytes. METHODS: Using 3-month-old New Zealand white rabbits, PRP was prepared from venous blood, and chondrocytes were cultured from knee joint cartilage and identified by staining for type II collagen and proteoglycan. The effects of PRP on chondrocyte viability were measured. The chondrocytes were divided into 5 groups: control, IL-1β, PRP (100-fold dilution), Dkk-1 (100ng/mL) and Dkk-1+PRP. The IL-1β, PRP, Dkk-1 and Dkk-1+PRP groups were treated with interleukin (IL)-1β (50μL, 10μg/mL) for24h. Chondrocyte morphology was observed by electron microscopy. Levels of carboxy terminal peptide (CTX-II) and cartilage oligomeric matrix protein (COMP) in culture media were measured by ELISA. Wnt-1, β-catenin and GSK-3β mRNA and protein expression were determined by RT-PCR and western blot respectively. RESULTS: PRP enhanced chondrocyte proliferation. Chondrocytes in the IL-1β group showed ultrastructural abnormalities that were less pronounced in the PRP, Dkk-1 and Dkk-1+PRP groups. CTX-II and COMP concentrations were higher in the IL-1β group than in the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). The IL-1β group had higher mRNA and protein Wnt1 and β-catenin levels and lower GSK-3β levels than the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). CONCLUSION: PRP may protect chondrocytes activated by IL-1β via inhibiting Wnt/β-catenin signaling.
Authors: Maria De Santis; Berardo Di Matteo; Emanuele Chisari; Gilberto Cincinelli; Peter Angele; Christian Lattermann; Giuseppe Filardo; Nicolò Danilo Vitale; Carlo Selmi; Elizaveta Kon Journal: Biomed Res Int Date: 2018-10-15 Impact factor: 3.411