Post-treatment of hyaluronan to decrease the apoptotic effects of carprofen in canine articular chondrocyte culture.

Korakot Nganvongpanit,Kittisak Buddhachat,Thippaporn Euppayo,Siriwan Ongchai,Siriwadee Chomdej,Puntita Siengdee

doi:10.7717/peerj.8355

Abstract

A major concern associated with the use of drugs is their adverse side effects. Specific examples of the drugs of concern include antibiotic agents and non-steroidal anti-inflammatory drugs. Despite the presence of a high degree of efficacy for specific conditions, these drugs may deteriorate the surrounding tissues that are exposed to them. Often, carprofen is used for joint inflammation; however, it may stimulate cartilage degradation which can then lead to osteoarthritis progression. In this study, hyaluronan was combined with carprofen treatment in three different applications (pre-treatment, co-treatment and post-treatment) on normal canine chondrocytes to determine whether Hyaluronan (HA) is capable of mitigating the degree of chondrotoxicity of carprofen. Our findings revealed that carprofen at IC20 (0.16 mg/mL) decreased viability and increased nitric oxide (NO) production. Importantly, carprofen induced the apoptosis of canine chondrocytes via the up-regulation of Bax, Casp3, Casp8, Casp9 and NOS2 as compared to the control group. Although the co-treatment of HA and carprofen appeared not to further alleviate the chondrotoxicity of carprofen due to the presence of a high number of apoptotic chondrocytes, post-treatment with HA (carprofen treatment for 24 h and then changed to HA for 24 h) resulted in a decrease in chondrocyte apoptosis by the down-regulation of Bax, Casp3, Casp8, Casp9, NOS2, along with NO production when compared with the treatment of carprofen for 48 h (P < 0.05). These results suggest that HA can be used as a therapeutic agent to mitigate the degree of chondrotoxicity of carprofen.

Highlights

Several drugs, such as local anesthetic drugs (10 mg/mL of lidocaine or 7.5 mg/mL of ropivacaine in human chondrocyte culture), corticosteroids
Chondrocyte apoptosis and nitric oxide (NO) production Morphological changes of the treated cells were observed under a phase contrast inverted microscope
This study found that the treatment at 0.16 mg/mL CAR either at 24 or 48 h could cause chondrocyte apoptosis to be observed via chromatin condensation, DNA fragmentation, and the apoptotic bodies that were presumably involved in the COX-2independent pathway at a molecular level by activating the up-regulation of some apoptotic genes (BAX, Caspase 3 (CASP3), Caspase 9 (CASP9)) and by inducing NO production

Summary

Introduction

Several drugs, such as local anesthetic drugs (10 mg/mL of lidocaine or 7.5 mg/mL of ropivacaine in human chondrocyte culture), corticosteroids 25 mg/mL of prednisolone or 7 mg/mL of betamethasone in human chondrocyte culture), cefazolin (2.0–3.33 mg/mL in canine chondrocyte culture) and fluoroquinolones (182 mg/mL of enrofloxacin or 295 mg/mL of marbofloxacin), have been shown to produce adverse effects on articular cartilage including apoptosis and the inhibition of growth of chondrocyte cells (Chrysis et al, 2005; Farkas et al, 2010; Nakazawa et al, 2002; Siengdee et al, 2016; Siengdee et al, 2017) This phenomenon can result in a loss of extra-cellular matrix formation and the progressive degradation of articular cartilage (Farkas et al, 2010). Non-steroidal anti-inflammatory drugs (NSAIDs), including indomethacin, ketorolac, diclofenac, piroxicam and celecoxib, caused proliferation suppression and the cell death of chondrocyte culture in vitro (Chang et al, 2006)

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