Effects of bromelain on cellular characteristics and expression of selected genes in canine in vitro chondrocyte culture

Siengdee, Puntita; Nganvongpanit, Korakot; Pothacharoen, Peraphan; Chomdej, Siriwadee; Mekchay, Supamit; Ongchai, Siriwan

doi:10.17221/3012-vetmed

Cited by 9 publications

(10 citation statements)

References 29 publications

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“…An enzymatic treatment was used to isolate cells for culture (Siengdee et al, 2010). Briefly, cartilage discs were washed three times with phosphate-buffered saline (PBS) containing 109 of a routine antibiotic dose of 2000 U/mL of penicillin, 2000 lg/mL of streptomycin, and 200 lg/mL of gentamicin.…”

Section: Sample Collection Cell Isolation and Cell Culturementioning

confidence: 99%

Two fluoroquinolones and their combinations with hyaluronan: comparison of effects on canine chondrocyte culture

Siengdee

Euppayo

Buddhachat

et al. 2016

Vet Pharm & Therapeutics

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Fluoroquinolones (FQs) are frequently used for septic arthritis. Increased antibacterial activity has been associated with mammalian cell cytotoxicity that may increase the risk of developing osteoarthritis. This study compared the direct effects of two different FQs, enrofloxacin (Enro) and marbofloxacin (Mar), on normal primary canine chondrocytes and inflammatory-stimulated chondrocytes, in addition to their administration in combination with hyaluronan (HA). Cell viability, cell apoptosis, s-GAG production, and expression patterns of inflammatory, extracellular matrix (ECM) component and protease genes were measured. Enro co-culturing with HA could modify s-GAG synthesis compared with the negative control group. Co-treatment with both FQs and HA significantly decreased cell viability and induced more total apoptotic cell death compared with the negative control and pre-IL-1β-stimulated group. Enro regulated IL-1β-stimulated cells to overexpress IL-1β, TNF, and MMP3, whereas Mar induced upregulation of PTGS2 and NFKB1 and enhanced the expression of ECM component genes HAS1, COL2A1, and ACAN as well as TIMP1 and MMP9. Simultaneous use of HA with Enro can effectively reduce the expression of IL-1β, TNF, and MMP3 in pre-IL-1β-stimulated chondrocytes. These results suggest the beneficial effects of HA in reducing the adverse effects of Enro treatment at the transcriptional level.

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Section: Sample Collection Cell Isolation and Cell Culturementioning

confidence: 99%

Two fluoroquinolones and their combinations with hyaluronan: comparison of effects on canine chondrocyte culture

Siengdee

Euppayo

Buddhachat

et al. 2016

Vet Pharm & Therapeutics

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“…The enzyme MMP-3 can cleave collagen, aggrecan, and link protein, while TIMPs inhibit the activity of MMPs. This study evaluates the expression of MMP-3 and TIMP-1 because these two genes are related, as previously described [36,37] . An increase in the level of the enzyme MMP-3 in comparison with TIMP-1 in the cartilage and the synovial membrane would explain the decrease in proteoglycan.…”

Section: Discussionmentioning

confidence: 99%

Köpeklerde Patellar Luksasyonda Kıkırdak Erozyonunun Prevalansı ve Etkilenmiş Eklemlerdeki Gen Ekspresyonu

Boonsri¹,

Pradit²,

Soontornvipart³

et al. 2016

Kafkas Univ Vet Fak Derg

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The objectives of this study were to assess the prevalence of cartilage erosion in small dogs with patellar luxation (PL), and related osteoarthritis (OA)-related gene expression. In Study 1, 71 dogs were examined to determine risk factors associated with PL, including breed, age, weight, sex, and affected joint. In Study 2, a total of 39 dogs were divided into four groups: normal articular cartilage in the stifle joint (G1; n=5); PL without cartilage erosion (G2; n=11); PL with cartilage erosion (G3; n=14); and OA in the stifle (G4; n=9). Articular cartilage and synovial membranes were collected during surgical operations to correct PL. Real-time PCR was used to quantify the expression levels of 11 OA-related genes, including AGG, COL2A1, HAS-1, with GAPDH used as a reference gene. From Study 1, it was found that the risk factors related with cartilage erosion lesion were age, sex, and PL grade (all variables showed P<0.05). From Study 2, it was demonstrated that PL with or without cartilage erosion expressed pro-inflammatory cytokines and enzymes; some biomolecules were up regulated (IL-1β, MMP-3, AGG, TIMP-1) but some were down regulated (COL2A1, . This expression was the difference between the articular cartilage and the synovial membrane; however, the expression of genes from PL with cartilage erosion was observed to be similar to that of OA. From our results, it can be concluded that PL can develop into secondary OA due to an increase of IL-1β in cartilage and synovial membrane. Keywords: Cartilage erosion, Dog, Gene expression, Patellar luxation Köpeklerde Patellar Luksasyonda Kıkırdak Erozyonunun Prevalansı ve Etkilenmiş Eklemlerdeki Gen Ekspresyonu ÖzetBu çalışmanın amacı patellar luksasyonlu (PL) küçük cüsseli köpeklerde kartilaj erozyonunun prevalansını ve osteoartritis (OA)-alakalı gen ekspresyonunu belirlemektedir. Birinci araştırmada; cins, yaş, cinsiyet ve infekte eklemi içeren PL ile ilgili risk faktörlerini belirlemek amacıyla 71 köpek incelendi. İkinci araştırmada toplam 39 köpek dört gruba ayrıldı; diz ekleminde normal artikular kartilaj (G1; n=5), kartilaj erozyon olmayan PL (G2; n=11), kartilaj erozyonlu PL (G3; n=14) ve dizde OA (G4; n=9). Cerrahi operasyon sırasında PL'u düzeltmek amacıyla artiküler kartilaj ve sinoviyal zarlar alındı. Toplam 11 adet OA ile ilgili genin (AGG, COL2A1,; referans gen olarak GAPDH kullanıldı) ekspresyon düzeylerini belirlemek amacıyla PCR tekniği uygulandı. Birinci araştırmanın sonucunda yaş, cinsiyet ve PL derecesi kartilaj erozyonu ile ilgili risk faktörleri olarak belirlendi (P<0.05). İkinci araştırmada kartilaj erozyonlu veya erozyon bulunmayan PL'lu köpeklerde pro-inflamatuar sitokinleri eksprese ettikleri, bazı biyomoleküllerin ekspresyonunu artırdıkları AGG, bazılarını ise azalttıkları (IL-1β, MMP-3, AGG, TIMP-1) gözlemlendi. Bu ekspresyon artiküler kartilaj ve sinoviyal zar için farklıydı. Ancak kartilaj erozyonlu PL'da genlerin ekspresyonu OA ile benzerlik göstermekteydi. Çalışma bulguları doğrultusunda kıkırdak ve sinoviyal zarlarda artan ...

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“…An ABI Prism 7000 sequence detection system (Applied Biosystems, Foster City, CA, USA) was used for quantitative analysis using SYBR Green JumpStart Taq ReadyMix (Sigma-Aldrich, St. Louis, MO, USA), incorporating dsDNA-specific fluorescent detection dye. Quantitative analyses of all transcriptions were performed in comparison with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) as an endogenous control and were run in separate wells [5]. PCR was performed using 2 μ L of each sample of cDNA and specific amplification primers.…”

Section: Methodsmentioning

confidence: 99%

“…After the end of the last cycle, a dissociation curve was generated by starting the fluorescence acquisition at 60°C and taking measurements at 7 s intervals until the temperature reached 95°C. Final quantitative analysis was done using the relative standard curve method, as in previous reports [5, 6]. Results were reported as the relative expression level compared to the calibrator cDNA after normalization of the transcript amount to the endogenous control.…”

Section: Methodsmentioning

confidence: 99%

Articular Cartilage Gene Expression after Coxofemoral Joint Luxation in the Dog

Nganvongpanit

Pradit

Chomdej

2013

Veterinary Medicine International

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This study examined the relationship between days of hip luxation and the expression of various mRNA. Twenty-six articular cartilages were used in the experiment: 3 samples were from normal dogs and 23 samples were collected from the femoral heads of hips that had been luxated for different lengths of time. Ten mRNA, including nonapoptotic genes (AGG, COL2A1, MMP-3, HAS-1, HAS-2, and TIMP-1) and apoptotic genes (BAX, BCL-2, CAS-3, and CAS-9), were studied for their expression using real-time PCR. We found very high correlation between expression level and luxation days (r 2 > 0.9) in COL2A1, MMP-3, HAS-1, HAS-2, TIMP-1, BAX, and CAS-9, while the others (AGG, BCL-2, and CAS-3) also showed high correlation (r 2 = 7–9). And we found a significant difference (P < 0.05) in the expression of transcripts depending on the number of luxation days. In conclusion, a delay in joint reduction may increase the chances of development of osteoarthritis.

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Effects of bromelain on cellular characteristics and expression of selected genes in canine in vitro chondrocyte culture

Cited by 9 publications

References 29 publications

Two fluoroquinolones and their combinations with hyaluronan: comparison of effects on canine chondrocyte culture

Two fluoroquinolones and their combinations with hyaluronan: comparison of effects on canine chondrocyte culture

Köpeklerde Patellar Luksasyonda Kıkırdak Erozyonunun Prevalansı ve Etkilenmiş Eklemlerdeki Gen Ekspresyonu

Articular Cartilage Gene Expression after Coxofemoral Joint Luxation in the Dog

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