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Effect of two different preparations of platelet-rich plasma on synoviocytes.

Assirelli E, Filardo G, Mariani E, Kon E, Roffi A, Vaccaro F, Marcacci M, Facchini A, Pulsatelli L - Knee Surg Sports Traumatol Arthrosc (2014)

Bottom Line: IL-1β, IL-8 and FGF-2 were significantly induced by L-PRP compared to both P-PRP and PPP; HGF was down-modulated by L-PRP versus both P-PRP and PPP, and an inverse dose-response influence was shown for all preparations.Expression level of TIMP-4 was lower in the presence of L-PRP compared with P-PRP.HA production and HAS gene expression did not seem to be modulated by PRP.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunorheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136, Bologna, Italy, elisa.assirelli@ior.it.

ABSTRACT

Purpose: To analyse the modifications induced by two different platelet-rich plasma (PRP) preparations on osteoarthritis (OA) synoviocytes, by documenting changes in gene expression of factors involved in joint physiopathology.

Methods: OA synoviocytes were cultured for 7 days in medium with different concentrations of either P-PRP (a pure platelet concentrate without leucocytes but with a limited number of platelets), L-PRP (a higher platelet concentrate containing leucocytes) or platelet-poor plasma (PPP). Gene expression of interleukin (IL)-1beta, IL-6, IL-8/CXCL8, tumour necrosis factor alpha, IL-10, IL-4, IL-13, metalloproteinase-13, tissue inhibitor of metalloproteinase (TIMP)-1, (TIMP)-3, (TIMP)-4, vascular endothelial growth factor, transforming growth factor beta1, fibroblast growth factor (FGF)-2, hepatocyte growth factor (HGF), hyaluronic acid (HA) synthases (HAS)-1, (HAS)-2, and (HAS)-3 was analysed by RT-PCR. HA production was determined in culture supernatants by ELISA.

Results: IL-1β, IL-8 and FGF-2 were significantly induced by L-PRP compared to both P-PRP and PPP; HGF was down-modulated by L-PRP versus both P-PRP and PPP, and an inverse dose-response influence was shown for all preparations. Expression level of TIMP-4 was lower in the presence of L-PRP compared with P-PRP. HA production and HAS gene expression did not seem to be modulated by PRP.

Conclusions: L-PRP is able to sustain the up-regulation of proinflammatory factors, (IL-1beta, IL-8 and FGF-2), together with a down-modulation of HGF and TIMP-4 expression, two factors that have been recognized as anti-catabolic mediators in cartilage, thus supporting the need to further optimize the PRP preparations to be applied in clinical practice.

No MeSH data available.


Related in: MedlinePlus

Gene expression analysis of interleukin (IL)-1beta, IL-8/CXCL8, IL-6, IL-10, tumour necrosis factor (TNF) alpha. Synovial fibroblasts were treated for 7 days with 5, 10, 20 % of L-PRP, P-PRP or PPP obtained from each subject (n = 7). Gene expression relative quantification was performed, and data are expressed as number of molecules *100,000 GAPDH. Boxes indicate the 25 and 75 % percentiles, whiskers indicate the minimum to maximum values, and bars indicate the median; p value significances are shown in tables beside each figure, as determined by General Linear Model statistical analysis and Kendall Tau correlation; ns not significant
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Fig1: Gene expression analysis of interleukin (IL)-1beta, IL-8/CXCL8, IL-6, IL-10, tumour necrosis factor (TNF) alpha. Synovial fibroblasts were treated for 7 days with 5, 10, 20 % of L-PRP, P-PRP or PPP obtained from each subject (n = 7). Gene expression relative quantification was performed, and data are expressed as number of molecules *100,000 GAPDH. Boxes indicate the 25 and 75 % percentiles, whiskers indicate the minimum to maximum values, and bars indicate the median; p value significances are shown in tables beside each figure, as determined by General Linear Model statistical analysis and Kendall Tau correlation; ns not significant

Mentions: Gene expression analysis indicated that IL-1β and IL-8/CXCL8 were significantly induced by L-PRP compared with both P-PRP and PPP (IL-1 β: p < 0.0005 L-PRP vs. P-PRP, p = 0.008 L-PRP vs. PPP; IL-8: p = 0.002 L-PRP vs. P-PRP, p = 0.008 L-PRP vs. PPP) (Fig. 1). Moreover, L-PRP also had a dose–response effect (Kendall Tau, IL-1β p = 0.013, IL-8 p = 0.011). Furthermore, if we consider the combined dose treatment effect on IL-1β gene expression, the dose–response curve showed a significantly different incremental trend induced by L-PRP from those of both P-PRP and PPP (p = 0.003) (Fig. 1). No significance between preparations was found on IL-6, IL-10 and TNF-α gene expression rate (Fig. 1), whereas IL-4 and IL-13 gene expression levels were not detectable in cultured synovial fibroblasts. Fig. 1


Effect of two different preparations of platelet-rich plasma on synoviocytes.

Assirelli E, Filardo G, Mariani E, Kon E, Roffi A, Vaccaro F, Marcacci M, Facchini A, Pulsatelli L - Knee Surg Sports Traumatol Arthrosc (2014)

Gene expression analysis of interleukin (IL)-1beta, IL-8/CXCL8, IL-6, IL-10, tumour necrosis factor (TNF) alpha. Synovial fibroblasts were treated for 7 days with 5, 10, 20 % of L-PRP, P-PRP or PPP obtained from each subject (n = 7). Gene expression relative quantification was performed, and data are expressed as number of molecules *100,000 GAPDH. Boxes indicate the 25 and 75 % percentiles, whiskers indicate the minimum to maximum values, and bars indicate the median; p value significances are shown in tables beside each figure, as determined by General Linear Model statistical analysis and Kendall Tau correlation; ns not significant
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4541703&req=5

Fig1: Gene expression analysis of interleukin (IL)-1beta, IL-8/CXCL8, IL-6, IL-10, tumour necrosis factor (TNF) alpha. Synovial fibroblasts were treated for 7 days with 5, 10, 20 % of L-PRP, P-PRP or PPP obtained from each subject (n = 7). Gene expression relative quantification was performed, and data are expressed as number of molecules *100,000 GAPDH. Boxes indicate the 25 and 75 % percentiles, whiskers indicate the minimum to maximum values, and bars indicate the median; p value significances are shown in tables beside each figure, as determined by General Linear Model statistical analysis and Kendall Tau correlation; ns not significant
Mentions: Gene expression analysis indicated that IL-1β and IL-8/CXCL8 were significantly induced by L-PRP compared with both P-PRP and PPP (IL-1 β: p < 0.0005 L-PRP vs. P-PRP, p = 0.008 L-PRP vs. PPP; IL-8: p = 0.002 L-PRP vs. P-PRP, p = 0.008 L-PRP vs. PPP) (Fig. 1). Moreover, L-PRP also had a dose–response effect (Kendall Tau, IL-1β p = 0.013, IL-8 p = 0.011). Furthermore, if we consider the combined dose treatment effect on IL-1β gene expression, the dose–response curve showed a significantly different incremental trend induced by L-PRP from those of both P-PRP and PPP (p = 0.003) (Fig. 1). No significance between preparations was found on IL-6, IL-10 and TNF-α gene expression rate (Fig. 1), whereas IL-4 and IL-13 gene expression levels were not detectable in cultured synovial fibroblasts. Fig. 1

Bottom Line: IL-1β, IL-8 and FGF-2 were significantly induced by L-PRP compared to both P-PRP and PPP; HGF was down-modulated by L-PRP versus both P-PRP and PPP, and an inverse dose-response influence was shown for all preparations.Expression level of TIMP-4 was lower in the presence of L-PRP compared with P-PRP.HA production and HAS gene expression did not seem to be modulated by PRP.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunorheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136, Bologna, Italy, elisa.assirelli@ior.it.

ABSTRACT

Purpose: To analyse the modifications induced by two different platelet-rich plasma (PRP) preparations on osteoarthritis (OA) synoviocytes, by documenting changes in gene expression of factors involved in joint physiopathology.

Methods: OA synoviocytes were cultured for 7 days in medium with different concentrations of either P-PRP (a pure platelet concentrate without leucocytes but with a limited number of platelets), L-PRP (a higher platelet concentrate containing leucocytes) or platelet-poor plasma (PPP). Gene expression of interleukin (IL)-1beta, IL-6, IL-8/CXCL8, tumour necrosis factor alpha, IL-10, IL-4, IL-13, metalloproteinase-13, tissue inhibitor of metalloproteinase (TIMP)-1, (TIMP)-3, (TIMP)-4, vascular endothelial growth factor, transforming growth factor beta1, fibroblast growth factor (FGF)-2, hepatocyte growth factor (HGF), hyaluronic acid (HA) synthases (HAS)-1, (HAS)-2, and (HAS)-3 was analysed by RT-PCR. HA production was determined in culture supernatants by ELISA.

Results: IL-1β, IL-8 and FGF-2 were significantly induced by L-PRP compared to both P-PRP and PPP; HGF was down-modulated by L-PRP versus both P-PRP and PPP, and an inverse dose-response influence was shown for all preparations. Expression level of TIMP-4 was lower in the presence of L-PRP compared with P-PRP. HA production and HAS gene expression did not seem to be modulated by PRP.

Conclusions: L-PRP is able to sustain the up-regulation of proinflammatory factors, (IL-1beta, IL-8 and FGF-2), together with a down-modulation of HGF and TIMP-4 expression, two factors that have been recognized as anti-catabolic mediators in cartilage, thus supporting the need to further optimize the PRP preparations to be applied in clinical practice.

No MeSH data available.


Related in: MedlinePlus