Analysis of the Anti-Inflammatory and Anti-Osteoarthritic Potential of Flonat Fast®, a Combination of Harpagophytum Procumbens DC. ex Meisn., Boswellia Serrata Roxb., Curcuma longa L., Bromelain and Escin (Aesculus hippocastanum), Evaluated in In Vitro Models of Inflammation Relevant to Osteoarthritis

Abstract

Osteoarthritis (OA) is a joint disease characterized by inflammation of the synovium, angiogenesis, cartilage degradation, and osteophyte formation. Harpagophytum Procumbens DC. ex Meisn., Boswellia Serrata Roxb., Curcuma longa L., Bromelain and Escin (Aesculus hippocastanum) are plants which extracts, together to Bromelain and Escin (Aesculus hippocastanum) are traditionally used in OA. However, their mechanistic role remains unclear. We aimed to investigate whether these bioactives alone or in combination (as in Flonat Fast®) can suppress TNF-α-induced inflammation, angiogenesis, and osteophyte formation using two cell models involved in OA: endothelial cells and monocytes. Each plant extract was evaluated for its polyphenol content, antioxidant activity, and toxicity. In endothelial cells and monocytes, expression of genes involved in OA was assessed, functional assays for inflammation and angiogenesis were performed, and impairment of reactive oxygen species production (ROS) was evaluated. Exposure of cells to the bioactives alone and in combination before cytokine stimulation resulted in differential counterregulation of several gene and protein expressions, including those for cyclooxygenases-2, metalloproteinase-9, transforming growth factor β1, and bone morphogenic protein-2. We demonstrated that these bioactives modulated monocyte adhesion to endothelial cells as well as cell migration and endothelial angiogenesis. Consistent with radical scavenging activity in the cell-free system, the bioactives curbed TNF-α-stimulated intracellular ROS production. We confirmed the potential anti-inflammatory and antiangiogenic effects of the combination of Harpagophytum procumbens, Boswellia, Curcuma, Bromelain, and Escin and provided new mechanistic evidence for their use in OA. However, further clinical studies are needed to evaluate the true clinical utility of these bioactives as supportive, preventive, and therapeutic agents.

Development of L-Lysine-Loaded PLGA Microparticles as a Controlled Release System for Angiogenesis Enhancement

“…Ades (Centers for Disease Control, Atlanta, USA) [ 47 ]. Cells were cultured in MCDB-131 medium supplemented with 15% FBS as previously described [ 48 ]. For cell treatments, the HMEC-1 (seeded at the density indicated in the legend figures) was cultured in MCDB-131 containing 15% FBS in the absence or presence of free LYS (90 and 180 µmol/L corresponding to the maximum cumulative release of LYS from MS/K-4 over 72 h), MS-4 (0.5 and 1.0 mg/mL), and MS/K-4 (0.5 and 1.0 mg/mL) for 24 to 72 h.…”

“…The migration of HMEC-1 was assessed using a wound-healing assay [ 48 ]. The HMEC-1 was seeded in 6-well plates and grown to confluence.…”

“…The total RNA was isolated using TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. RNA was retrotranscribed to cDNA as previously described [ 48 ]. Real-time PCR (qPCR) analyses were performed using the CFX96 Touch Real-Time PCR Detection System and software (Bio-Rad Laboratories, Segrate, Italy).…”

“…Real-time PCR (qPCR) analyses were performed using the CFX96 Touch Real-Time PCR Detection System and software (Bio-Rad Laboratories, Segrate, Italy). All reactions were carried out in a total volume of 25 μL with 50 ng of cDNA, 0.3 pmol/L of the primer pair, and 12.5 μL 2× SYBR Green PCR master mix (Bio-Rad Laboratories, Segrate, Italy) under conditions that were previously described [ 48 ]. The quantifications were performed using the comparative critical threshold method (ΔΔCT), and the GAPDH and 18S genes were used as internal controls for normalization.…”

Fitoterapia en Osteoartritis: Eficacia y mecanismos implicados

“…En el análisis de Flonat Fast, una combinación de Harpagophytum procumbens DC. ex Meisn., Boswellia Serrata Roxb., Cúrcuma longa L., Bromelina y Escina (Aesculus hippocastanum) tiene un potencial para reducir algunas características patogenéticas de la progresión de la enfermedad de OA, incluida la diapédesis de monocitos y la adhesión al endotelio, la angiogénesis proinflamatoria y la síntesis y liberación de mediadores proinflamatorios y del dolor mostrando un mejor efecto de manera combinada que monoterapia estos extractos fitoterápicos (36,63).…”