Research ArticleHEALTH AND MEDICINE

Systemic anti-inflammatory therapy aided by double-headed nanoparticles in a canine model of acute intraocular inflammation

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Science Advances  26 Aug 2020:
Vol. 6, no. 35, eabb7878
DOI: 10.1126/sciadv.abb7878
  • Fig. 1 Synthesis and characterization of PLGA-GA2 and their CUR encapsulated nanoparticles (PLGA-GA2-CUR).

    (A) Schematic illustration of synthesis of PLGA-GA2. (B) Dynamic light scattering size distribution of PLGA-GA2-CUR with an insert depicting model particle. (C) Scanning electron microscopy photomicrographs of PLGA-GA2-CUR. TFA, trifluoroacetic acid; DIEA, N,N-diisopropylethylamine; TREN, Tris(2-aminoethyl)amine; EDC, 1-ethyl-3- (dimethylaminopropyl) carbodiimide.

  • Fig. 2 Target tissue distribution of oral PLGA-GA2-CUR in healthy dogs and anti-inflammatory effects of oral PLGA-GA2-CUR in a canine model of acute intraocular inflammation.

    (A) Aqueous humor levels of CUR following a single oral dose of PLGA-GA2-CUR. (B to D) Following intracameral injection of lens protein at t = 0 hours, eyes were serially evaluated using the SPOTS system, including measurements of (B) aqueous flare, (C) pupillary light reflex, (D) conjunctival swelling. Statistical significance of oral PLGA-GA2-CUR administration was compared with treatment with oral carprofen and untreated controls, as determined by a two-way ANOVA. Statistical significance is denoted by *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

  • Fig. 3 Anti-inflammatory effects of topical PLGA-GA2-CUR in a canine model of acute intraocular inflammation.

    (A to C) Following intracameral injection of lens protein at t = 0 hours, eyes were serially evaluated using the SPOTS system, including measurements of (A) aqueous flare, (B) pupillary light reflex, (C) conjunctival swelling. Statistical significance of topical PLA-GA2-CUR administration was compared to treatment with topical prednisolone acetate and untreated controls, as determined by a two-way ANOVA. Statistical significance is denoted by *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

  • Table 1 Summary of hematology values in healthy dogs before and after oral PLGA-GA2-CUR administration.

    WBC, white blood cells; Abs, absolute; Rel, relative; RBC, red blood cell; HGB, hemoglobin; HCT, hematocrit; MCV, mean corpuscular volume; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; RDW, red cell distribution width; PLT, platelet; MPV, mean platelet volume; PCT, procalcitonin; Retic, reticulocyte; Retic abs count, absolute reticulocyte count.

    CBC whole bloodBefore dosing24 hours after dosing PLGA-GA2-CUR7 days after dosing PLGA-GA2-CURReference range
    Total WBC (K/μl)13.46 ± 310.79 ± 2.3511.5 ± 1.366.0–17.0
    Abs neutrophils (K/μl)8.93 ± 2.946.00 ± 1.46.98 ± 1.533.0–11.5
    Abs lymphocytes (K/μl)3.7 ± 0.463.95 ± 1.043.33 ± 0.811.0–4.8
    Abs monocytes (K/μl)0.57 ± 0.420.6 ± 0.290.9 ± 0.220.15–1.35
    Abs eosinophils (K/μl)0.27 ± 0.060.3 ± 0.10.25 ± 0.170.10–1.25
    Rel neutrophils (%)65.33 ± 6.8155.5 ± 1.2960.5 ± 8.58
    Rel lymphocytes (%)29 ± 9.5436.25 ± 2.8729.25 ± 7.63
    Rel monocytes (%)4.00 ± 3.006.25 ± 3.778.00 ± 2.00
    Rel eosinophils (%)3.67 ± 0.583.46 ± 1.652.25 ± 1.26
    RBC (M/μl)5.94 ± 0.278.08 ± 3.766.54 ± 0.275.5–8.5
    HGB (g/dl)10.6 ± 4.8120.35 ± 12.7715.13 ± 0.9312.0–18.0
    HCT (%)28.1 ± 13.946.9 ± 13.9743.55 ± 0.8632–55
    MCV (fl)51.17 ± 16.6954.23 ± 20.6966.68 ± 3.3660–77
    MCH (pg)35.8 ± 23.1325.6 ± 6.1723.15 ± 0.9319.5–24.5
    MCHC (g/dl)32.53 ± 7.6829.73 ± 10.4934.73 ± 0.3832–36
    RDW (%)22.63 ± 13.2215.15 ± 1.514.63 ± 01.5911.9–14.9
    PLT (K/μl)445.4 ± 66.72444.33 ± 21.95521.25 ± 79.28200–500
    MPV (fl)9.00 ± 3.897.46 ± 5.139.9 ± 1.757.9–16.2
    PCT (%)3.27 ± 4.70.53 ± 0.20.52 ± 0.150.12–0.43
    Retic (%)0.66 ± 0.340.94 ± 0.611.39 ± 0.84
    Retic abs count (K/μl)43.46 ± 14.3041.4 ± 22.750.66 ± 11.4210–80
  • Table 2 Summary of serum biochemistry values in healthy dogs before and after oral PLGA-GA2-CUR administration.

    GA2-CUR administration. A/G ratio, albumin-to-globulin ratio; BUN, blood urea nitrogen; ALKP, alkaline phosphatase; AST, aspartate aminotransaminase; ALT, alanine aminotransaminase; GGT, gamma-glutamyl transferase; GLDH, glutamate dehydrogenase.

    Chemistry profileBefore dosing24 hours after dosing
    PLGA-GA2-CUR
    7 days after dosing
    PLGA-GA2-CUR
    Reference range
    Total protein (g/dl)5.33 ± 0.155.03 ± 0.105.13 ± 0.105.6–7.9
    Albumin (g/dl)2.98 ± 0.172.93 ± 0.053.05 ± 0.062.8–4.3
    Globulins (g/dl)2.35 ± 0.192.10 ± 0.082.08 ± 0.051.8–4.2
    A/G ratio1.30 ± 0.181.40 ± 0.081.48 ± 0.050.7–2.5
    Calcium (mg/dl)10.75 ± 0.3410.48 ± 0.2610.63 ± 0.137.2–12.8
    Phosphorus (mg/dl)7.45 ± 0.706.93 ± ±0.547.03 ± 0.212.3–6.5
    Glucose (mg/dl)73.00 ± 10.30100.00 ± 6.88106.25 ± 6.7060–120
    BUN (mg/dl)14.25 ± 1.2612.75 ± 1.0510.25 ± 1.508–30
    Creatinine (mg/dl)0.53 ± 0.050.58 ± 0.100.63 ± 0.050.5–1.4
    Bilirubin–total (mg/dl)0.15 ± 0.100.20 ± 0.080.15 ± 0.060.1–0.4
    ALKP (U/liter)108.50 ± 25.72107.25 ± 16.66103.50 ± 16.6012–122
    AST (U/liter)46.25 ± 4.7935.25 ± 1.5040.50 ± 2.6513–52
    ALT (U/liter)34.50 ± 3.7032.50 ± 5.3237.75v7.4113–79
    GGT (U/liter)4.25 ± 1.262.50 ± 0.713.25 ± 0.500–10
    GLDH (U/liter)8.50 ± 1.917.75 ± 0.966.50 ± 1.290–14
    Amylase (U/liter)592.00 ± 93.03590.00 ± 115.86605.50 ± 47.76200–953
    Cholesterol (mg/dl)172.75 ± 18.52164.50 ± 20.86160.50 ± 17.71124–335
    Sodium (mEq/liter)146.75 ± 1.71145.75 ± 0.50146.00 ± 1.63141–156
    Potassium (mEq/liter)5.18 ± 0.384.75 ± 0.131.93 ± 0.133.8–5.5
    Chloride (mEq/liter)107.50v0.58109.50 ± 0.58109.50 ± 0.58109–124
    Sodium-to-potassium ratio28.45 ± 1.8330.70 ± 0.8629.65 ± 0.82
  • Table 3 P values of the test H0:β¯3=0 using model (2) and H0:β2=0 using model (3) for all three comparisons under oral administration.

    Three comparisons under oral administration.

    Pairwise comparisonH0:β¯3=0H0:β2=0
    Untreated versus carprofen0.138270.014
    Untreated versus PLGA-GA2-CUR0.870490.00483
    Carprofen versus PLGA-GA2-CUR0.153260.7270
  • Table 4 P values of the test H0:β¯3=0 using model (2) and H0:β2=0 using model (3) for all three comparisons under topical administration.

    Pairwise comparisonH0:β¯3=0H0:β2=0
    Untreated versus prednisolone
    acetate
    0.110370.00438
    Untreated versus PLGA-GA2-CUR0.557480.00864
    Prednisolone acetate versus
    PLGA-GA2-CUR
    0.185370.506844

Supplementary Materials

  • Supplementary Materials

    Systemic anti-inflammatory therapy aided by double-headed nanoparticles in a canine model of acute intraocular inflammation

    R. Ganugula, M. Arora, M. A. Lepiz, Y. Niu, B. K. Mallick, S. C. Pflugfelder, E. M. Scott, M. N. V. Ravi Kumar

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