title: Intra-articular interleukin-1 receptor antagonist (IL1-ra) microspheres for posttraumatic osteoarthritis: in vitro biological activity and in vivo disease modifying effect.
abstract: BACKGROUND
  Interleukin-1 receptor antagonist (IL-1 ra) can be disease-modifying in posttraumatic osteoarthritis (PTOA). One limitation is its short joint residence time. We hypothesized that IL-1 ra encapsulation in poly (lactide-co-glycolide) (PLGA) microspheres reduces IL-1 ra systemic absorption and provides an enhanced anti-PTOA effect.
  
  
  METHODS
  IL-1 ra release kinetics and biological activity: IL-1 ra encapsulation into PLGA microsphere was performed using double emulsion solvent extraction. Lyophilized PLGA IL-1 ra microspheres were resuspended in PBS and supernatant IL-1 ra concentrations were assayed. The biological activity of IL-1 ra from PLGA IL-1 ra microspheres was performed using IL-1 induced lymphocyte proliferation and bovine articular cartilage degradation assays. Systemic absorption of IL-1 ra following intra-articular (IA) injection of PLGA IL-1 ra or IL-1 ra: At 1, 3, 6, 12 and 24 h following injection of 50 μl PLGA IL-1 ra (n = 6) or IL-1 ra (n = 6), serum samples were collected and IL-1 ra concentrations were determined. Anterior cruciate ligament transection (ACLT) and IA dosing: ACLT was performed in 8-10 week old male Lewis rats (n = 42). PBS (50 μl; n = 9), IL-1 ra (50 μl; 5 mg/ml; n = 13), PLGA IL-1 ra (50 μl; equivalent to 5 mg/ml IL-1 ra; n = 14) or PLGA particles (50 μl; n = 6) treatments were performed on days 7, 14, 21 and 28 following ACLT. Cartilage and synovial histopathology: On day 35, animal ACLT joints were harvested and tibial cartilage and synovial histopathology scoring was performed.
  
  
  RESULTS
  Percent IL-1 ra content in the supernatant at 6 h was 13.44 ± 9.27 % compared to 34.16 ± 12.04 %, 47.89 ± 12.71 %, 57.14 ± 11.71 %, and 93.90 ± 8.50 % at 12, 24, 48 and 72 h, respectively. PLGA IL-1 ra inhibited lymphocyte proliferation and cartilage degradation similar to IL-1 ra. Serum IL-1 ra levels were significantly lower at 1, 3, and 6 h following PLGA IL-1 ra injection compared to IL-1 ra. Cartilage and synovial histopathology scores were significantly lower in the PLGA IL-1 ra group compared to PBS and PLGA groups (p < 0.001).
  
  
  CONCLUSIONS
  IL-1 ra encapsulation in PLGA microspheres is feasible with no alteration to IL-1 ra biological activity. PLGA IL-1 ra exhibited an enhanced disease-modifying effect in a PTOA model compared to similarly dosed IL-1 ra.
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy-Boston, MCPHS University, Boston, MA, USA. elsaid@chapman.edu.|Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, 9401 Jeronimo Road, Irvine, CA, 92618, USA. elsaid@chapman.edu.
authors.forename: Khaled A
authors.initials: KA
authors.lastname: Elsaid
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy-Boston, MCPHS University, Boston, MA, USA.
authors.forename: Anand
authors.initials: A
authors.lastname: Ubhe
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy-Boston, MCPHS University, Boston, MA, USA.
authors.forename: Ziyad
authors.initials: Z
authors.lastname: Shaman
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy-Boston, MCPHS University, Boston, MA, USA.
authors.forename: Gerard
authors.initials: G
authors.lastname: D'Souza
keywords: Bioactivity of IL1ra; IL1 ra microspheres; Interleukin-1 receptor antagonist; Posttraumatic osteoarthritis