Novel formulations ameliorate osteoarthritis in rats by inhibiting inflammation and oxidative stress

Abstract

The study tested new oral plant-based formulations (F) on rats with monosodium iodoacetate (MIA)-induced osteoarthritis, measuring inflammation, antioxidant levels, paw size, stride, and analyzing knee joint images. Fifty-six female Sprague Dawley rats were allocated into 8 groups: (1) Control, (2) MIA (OA induced with MIA), (3) MIA + F1 [curcuminoids+gingerols+acetyl-11-keto-β boswellic acid (AKBA)], (4) MIA + F2 (curcuminoids+Withania glycosides+AKBA), (5) MIA + F3 (curcuminoids+total withanolides+AKBA), (6) MIA + F4 (curcuminoids, AKBA), (7) MIA + UCII (type II collagen), and (8) MIA + GCHON (Glucosamine Chondroitin). Treatments F1 to F4 reduced right joint diameter and improved stride length and paw area in OA rats. Despite improvements with treatments F1 to F4, there was no significant difference between these groups (p > .05). In OA animals, F1 to F4 treatments decreased MDA levels and increased antioxidant enzymes activities (p < .001). This was done by reducing levels of inflammatory markers and enzymes like IL-1β, IL-6, MMP-8, TNF-α, CRP, COMP, and LOX-5, while increasing the anti-inflammatory cytokine IL-10. In conclusion, these plant-based treatments significantly reduced osteoarthritis severity, slowed disease progression by reducing inflammation, and protected joints from damage, showing a protective effect in rats with induced osteoarthritis, likely due to their anti-inflammatory and antioxidant properties.

Keywords: anti‐inflammatory; inflammation; osteoarthritis; oxidative stress; plant‐based formulations.

FIGURE 1

Effects of different OAHT formulations on rat knee swelling (A) and knee joint diameter (B) associated with monosodium iodoacetate (MIA)‐induced OA. The bars and error lines point out the mean and standard deviation. ANOVA and Tukey's post hoc test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the groups (p < .05). F1:Curcuminoids+Gingerols+acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4:Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.

FIGURE 2

Effects of different OAHT formulations on rat paw area (A), paw width (B), and stride length (C) in monosodium iodoacetate (MIA)‐induced OA. Representative measures are shown (D). The bars and error lines point out the mean and standard deviation. ANOVA and Tukey's post hoc test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the groups (p < .05). F1: Curcuminoids+Gingerols+acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4:Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.

FIGURE 3

Effects of different OAHT formulations on the rat knee joint with monosodium iodoacetate (MIA)‐induced OA. Representative radiographic images are shown (A). Mean values of Kellgren–Lawrence scores are demonstrated (B) with standard deviation. Kruskal–Wallis and uncorrected Dunn's multiple comparisons test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the groups (p < .05). F1:Curcuminoids+Gingerols+ acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4:Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.

FIGURE 4

Effects of different OAHT formulations on rat knee joint histopathology associated with monosodium iodoacetate (MIA)‐induced OA. Representative hematoxylin–eosin stained joint cartilages are shown (A). Mean values of structure (B), cells (C), tidemark (D), and total Mankin scores (E) are demonstrated with standard deviation. Kruskal–Wallis and uncorrected Dunn's multiple comparisons test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the groups (p < .05). F1: Curcuminoids+Gingerols+ acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4:Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.

FIGURE 5

Effects of different OAHT formulations on serum IL‐β (A), IL‐6 (B), TNF‐α (C), COMP (D), and CRP (E) levels in rats with monosodium iodoacetate (MIA)‐induced OA. The bars and error lines point out the mean and standard deviation. ANOVA and Tukey's post hoc test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the study groups (p < .05). IL‐1β, interleukin‐1β; IL‐6, interleukin‐6; TNF‐ α, tumor necrosis factor α; COMP, cartilage oligomeric matrix protein; CRP, C‐reactive protein. F1: Curcuminoids+Gingerols+ acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4:Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.

FIGURE 6

Effects of different OAHT formulations on serum MDA (A), SOD (B), GSH‐Px (C), and CAT (D) levels in rats with monosodium iodoacetate (MIA)‐induced OA. The bars and error lines point out the mean and standard deviation. ANOVA and Tukey's post hoc test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the groups (p < .05). MDA, malondialdehyde; SOD, superoxide dismutase; GSH‐Px, glutathione peroxidase; CAT, catalase. F1: Curcuminoids+Gingerols+ acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4:Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.

FIGURE 7

Effects of different joint health formulations on knee joint protein expression of IL‐1β (A), IL‐6 (B), IL‐10 (C), TNF‐α (D), COMP (E), and MMP‐8 (F), levels in monosodium iodoacetate (MIA)‐induced OA rats. The densitometric analysis of the relative intensity according to the control group of the Western blot bands was performed with β‐Actin normalization to ensure equal protein loading. Blots were repeated at least three times (n = 3), and a representative blot is shown. The bars and error lines point out the mean and standard deviation. ANOVA and Tukey's post hoc test were performed for statistical comparison. Different letters (a, b, and c) above the bars indicate statistical differences among the groups (p < .05). IL‐1β, interleukin‐1β; IL‐6, interleukin‐6; IL‐10, interleukin‐10; TNF‐ α, tumor necrosis factor α; COMP, cartilage oligomeric matrix protein; MMP‐8, matrix metalloproteinase‐8. F1: Curcuminoids+Gingerols+acetyl‐11‐keto‐β boswellic acid (AKBA); F2: Curcuminoids+Withania Glycosides+ AKBA; F3: Curcuminoids+Total Withanolides+AKBA; F4: Curcuminoids+AKBA; UCII: Undenatured type II collagen; G + CHON: Glucosamine Hydrochloride, Chondroitin Sulfate, Hyaluronic Acid, Calcium Fructoborate.