CAVALOR ARTITEC – The result of an extensive 7-year research program

THE EFFICACY AND ESPECIALLY THE SYNERGISTIC POTENTIAL of the individual raw materials and botanical ingredients on inflammation, cartilage protection and repair has been studied over the last 7 years. These findings have led to the final formulation of Cavalor ArtiTec.

RESEARCH PROGRAM CAVALOR ARTITEC PERFORMED BY UNIVERSITY OF PRINCE EDWARD ISLAND, CANADA

  • PHASE 1 – IN VITRO – Screening and selection of individual ingredients for their anti-inflammatory effect
    • Osteoarthritis model
  • PHASE II-I – IN VIVO Optimizing the product formulation
    • Screening the efficacy of various multiherbal formulations for the additive effects in an inflammation model
  • PHASE II-II – IN VITRO – Optimizing the product formulation
    • Product optimization: Evaluation of ingredient combinations in a joint inflammation model
  • FINAL TEST FORMULA (IN VIVO) – FACULTY OF VETERINARY MEDICINE, GHENT UNIVERSITY
  • PHASE III
    • Testing the preventive effects of CAVALOR ARTITEC on experimentally induced acute synovitis in horses (Published in Equine Veterinary Journal, 2016)
  • PHASE IVCAVALOR ARTITEC Clinical Trial
    • Preliminary Field trial performed by Belgian veterinarians

CAVALOR ARTITEC > FOR JOINTS THAT PERFORM > PHASE 1

Cavalor ArtiTec Research > Effect of herbal extract on monocytes production tumor necrosis factor (TNF) – IN VITRO MODEL – Prince Edward Island –  University of Canada

> IN VITRO MODEL

  • The purpose of this study is to provide insight into the molecular mechanism by which Cavalor ArtiTec may reduce articular injuring during osteoarthritis.

> PROTOCOL

  • Monocytes (THP-1) were stimulated with IL-1B and Lipopolysaccharide (LPS). Gene expression of Tumor Necrosis Factor (TNF) was analyzed by ‘reverse transcription polymerase chain reaction’.

> IN A FIRST PART, THE EFFECT OF SEPARATE INGREDIENTS WAS MEASURED

  • Different ingredients reduce TNF expression with IL-1B stimulation other ingredients had an effect on LPS stimulation.

PHASE II-I

SCREENING AND SELECTION OF INDIVIDUAL INGREDIENTS FOR THEIR ANTI-INFLAMMATORY EFFECT

A first phase of research was performed by ‘University of Prince Edward Island’. Some of the investigated ingredients were able to downregulate the expression of Tumor Necrosis Factor (TNF) of monocytes stimulated by Lipopolysaccharide (LPS). On the other hand, in the same research phase a number of ingredients were identified that potentiated LPS-induced TNF expression.

TNF GENE EXPRESSION > BY HUMAN MONOCYTES STIMULATED WITH LPS 

Monocytes are a type of white blood cell, or leukocyte. They are the largest type of leukocyte, can differentiate into macrophages and can acquire some features of dendritic cells. As a part of the vertebrate innate immune system, monocytes also influence the process of adaptive immunity.

Optimizing the ArtiTec Product Formulation

PHASE II-I: SCREENING THE EFFICACY OF VARIOUS MULTIHERBAL FORMULATIONS FOR THE ADDITIVE EFFECTS IN AN INFLAMMATION MODEL

Combining botanical ingredients is a widely applied practice to compose effective products. The final result is believed to be of similar or better potency as that of a single herb. However, there is a lack of studies executed in animals that have provided evidence regarding this “synergistic effect” of herbs. Cavalor initiated a study to demonstrate the additive effect of herbal mixtures and to characterize their range of anti-inflammatory activity. The advantage of using suboptimal efficacious dose levels of each botanical ingredient is reducing the risk of potential toxicities associated with the usage of single herbs.

Various combinations of ingredients were compared in a validated (inflammation) model that assessed the potential to reduce (acute) inflammation following a single oral administration. The applied formulation was composed of the ingredient combination A, B, C, D and E.

While maintaining a total dose of the extract fixed, additional herb extracts were added to demonstrate the additive effect of the combination. For example, the amount of ingredient A was 250 mg and of A+B+C+D (62.5 +62.5 + 62.5 + 62.5) was also 250 mg. The results of this study were used to define and evaluate candidate product formulations in Phase II-II.

PRODUCT FORMULATION

PHASE II-II: EVALUATION OF 7 DIFFERENT INGREDIENT COMBINATIONS – RAW MATERIALS & MULTIHERBAL FORMULATIONS IN A JOINT INFLAMMATION MODEL

In the follow-up of the results of previous studies, 7 candidate formulations were evaluated (in vitro) to further unlock the mechanism of action of these formulations and their efficacy. The influence on various biomarkers related to joint health and homeostasis were evaluated using primary human chondrocytes isolated from knee joints in various in vitro models.

This study has led to the development of the final candidate formulation for evaluation in a combined mechanistic and efficacy horse specific joint inflammation model (Phase III).

> Primary human chondrocytes isolated from human knee joints during arthroscopic surgery

> Chondrocytes pretreated with herbs/combinations (25 μg/ml final concentration of each herb) for 24 hr

CAVALOR ARTITEC (C6) PROTECTS THE CARTILAGE AGAINST INFLAMMATION

DOWNREGULATE EXPRESSION FAS RECEPTOR

CAVALOR ARTITEC (C6) STIMULATES THE REPAIR OF THE CARTILAGE TISSUE

UPREGULATE EXPRESSION BMP-5

PHASE III > Cavalor ArtiTec Research – EXPERIMENTAL IN VIVO STUDY (HORSES)

ATL-CH STUDY

GHENT UNIVERSITY (ELINE VAN DE WATER, MAARTEN OOSTERLINCK, FREDERIK PILLE)

> PROTOCOL

24 healthy and clinically sound Standardbred horses

  • 24 healthy and clinically sound Standardbred horses
  • 4 treatment groups
  • including a positive (meloxicam) and negative (placebo) control group

> INTRA-ARTICULAR CHALLENGE

  • with 0.5 ng LPS of coli

> EVALUATION OF TREATMENTS

  • synovial fluid (SF) analysis: cytology, total protein, biomarkers lameness evaluation: clinical lameness score, pressure plate analysis

> REMARK

  • joint inflammation is accompanied by cellular and vascular changes resulting in an influx of inflammatory cells and serum proteins, as illustrated by the TNCC and TP values, and an up-regulation of specific mediators such as PGE2

TESTING THE PREVENTITIVE EFFECT OF CAVALOR ARTITEC ON EXPERIMENTALLY INDUCED ACUTE SYNOVITIS IN HORSES

SYNOVIAL FLUID ANALYSIS

> LEFT: synovial fluid sample before injection of LPS, presenting clear, straw-colored fluid with normal viscosity.

> RIGHT: synovial fluid sample 8 hours after injection of LPS, presenting brownish, turbid fluid with decreased viscosity, as a result of acute inflammation.

> TOTAL NUCLEATED CELL COUNTS (TNCC)

> TOTAL PROTEIN (TP)

> BIOMARKERS

  • Prostaglandin E2 (PGE2)

> PRESSURE PLATE ANALYSIS

  • Peak vertical force (PVF)

CONCLUSION (PHASE III)

  • Cavalor ArtiTec significantly reduces he inflammation after experimentally. Induced synovitis (LPS) and reduces lameness compared to placebo (PVF)

FIELD TRIAL – Pilot Study

INTRODUCTION

Cavalor ArtiTec Liquid was evaluated in a multi-practitioners field study for the treatment of horses with mild foot lameness with or without relevant imaging findings.

INCLUSION & EXCLUSION CRITERIA

Only horses with a lameness of more than 3 months duration and localized to the foot by a positive digital palmar nerve block were included.

Cavalor ArtiTec Liquid was administered at a dose of 90ml/day for 1 month. During the supplementation period, no concurrent treatments were accepted and for local infiltrations, a washout period of 6 weeks was required before entering the study. Shoeing adaptations were accepted until 4 weeks before entering the study. During the supplementation period, horses had to remain active at their usual level. At the end of the supplementation period, information about the palatability and the efficacy of the product was obtained by riders’ questionnaire and the horses were re-evaluated by the equine practitioner involved.

RESULTS

Thirty-one cases met the inclusion criteria and were enrolled in the study. Most of them were show jumpers performing at a national or international level. Apart from one case, the palatability of Cavalor ArtiTec Liquid was evaluated to be good or excellent. The efficacy of the product was rated similarly by both riders and equine practitioners: in 8/31 (26%) horses, there was no relevant change in observed lameness, whereas in 23/31 (74%) horses, lameness improved moderately (8/31; 26 %) or substantially (15/31; 48%).

CONCLUSION (PHASE IV)

> In conclusion, the field study supports the results obtained during the experimental in vivo study performed by Ghent University. Cavalor ArtiTec has received positive feedback from riders, as well as equine practitioners in the treatment of foot lameness and is therefore considered a real asset to the portfolio of therapeutics of equine practitioners. 

GENERAL CONCLUSION

Low-grade inflammation is common in joints that perform. However, joint inflammation initiates a cascade of catabolic reactions that gradually degrade the cartilage and, in the end, may result in lameness.

During an extensive research program, Cavalor has designed step by step a multi-ingredient nutraceutical that is able to downregulate synovial inflammation, hence protecting the joints of equine athletes. The anti-inflammatory effect of Cavalor ArtiTec has been documented by both in vitro and in vivo experiments and the overall efficacy in the treatment of lameness is supported by the results of a clinical pilot study.

In conclusion, Cavalor ArtiTec is a complete multi-ingredient nutraceutical with proven efficacy for optimizing joint health in equine athletes, hence being an asset to the portfolio of therapeutics used by equine practitioners.