Gene expression of bone-associated cytokines in MG63 osteoblast-like cells incubated with acrylic bone cement extracts in minimum essential medium.
This study examined the effects of in vitro challenge with four polymerized acrylic bone cements (Sulfix-60, CMW 1, CMW 2, and CMW 3) on the expression of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and transforming growth factor-beta1 (TGF-beta1) mRNAs in the osteoblast-like cell line MG63. The extracts of the cements in minimal essential medium (MEM) were tested following 1-h and 7-day curing. A semi-quantitative analysis of the cytokine-specific mRNAs was carried out by agarose gel densitometry and expression was compared with the GAPDH housekeeping gene. The ratio between cytokine gene expression and GAPDH expression was calculated. The mRNA specific for the bone-resorbing cytokines IL-1beta and IL-6 was low in basal conditions. IL-1beta mRNA increased only after incubation with the extract of CMW 1 following 1-h curing. The mRNA specific for the bone-resorbing cytokine IL-6 also increased after contact with CMW 1 at both curing times. Sulfix-60 and CMW 3 following 7-day curing, but not after 1 h, induced higher levels of IL-6 mRNA than the control. CMW 2 after 1-h curing constantly determined the expression of IL-6 mRNA, but at low levels. The mRNA specific for TGF-beta1 was also expressed by the MG63 osteoblast-like cells in basal conditions. The levels increased after contact with Sulfix-60 after 7-day curing and with CMW 1 after 1-h curing. CMW 2 after 7-day curing decreased TGF-beta1 mRNA. In conclusion, the highest expression of the cytokines IL-1beta, IL-6, and TGF-beta1 mRNA was determined by CMW 1. If the results are confirmed in vivo, the increased expression of the osteolytic cytokines induced by the bone cement might result in loosening of the prosthesis, even with all the restrictions of an in vitro study on continuous cell lines.
Cenni
E
E
Laboratorio di Fisiopatologia degli Impianti Ortopedici, Istituti Ortopedici Rizzoli, via di Barbiano 1/10, 40136 Bologna, Italy. elisabetta.cenni@ior.it
Granchi
D
D
Ciapetti
G
G
Savarino
L
L
Corradini
A
A
Vancini
M
M
Giunti
A
A
D001843:Bone Cements; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006133:Growth Substances / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis / Q000235:genetics; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D019904:Polymethyl Methacrylate; D011475:Prosthesis Failure; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics
D001843:Bone Cements; D006133:Growth Substances; D007375:Interleukin-1; D015850:Interleukin-6; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D019904:Polymethyl Methacrylate