Transdermal cannabidiol reduces inflammation and pain-related behaviours in a rat model of arthritis.

BACKGROUND: Current arthritis treatments often have side-effects attributable to active compounds as well as route of administration. Cannabidiol (CBD) attenuates inflammation and pain without side-effects, but CBD is hydrophobic and has poor oral bioavailability. Topical drug application avoids gastrointestinal administration, first pass metabolism, providing more constant plasma levels. METHODS: This study examined efficacy of transdermal CBD for reduction in inflammation and pain, assessing any adverse effects in a rat complete Freund's adjuvant-induced monoarthritic knee joint model. CBD gels (0.6, 3.1, 6.2 or 62.3 mg/day) were applied for 4 consecutive days after arthritis induction. Joint circumference and immune cell invasion in histological sections were measured to indicate level of inflammation. Paw withdrawal latency (PWL) in response to noxious heat stimulation determined nociceptive sensitization, and exploratory behaviour ascertained animal's activity level. RESULTS: Measurement of plasma CBD concentration provided by transdermal absorption revealed linearity with 0.6-6.2 mg/day doses. Transdermal CBD gel significantly reduced joint swelling, limb posture scores as a rating of spontaneous pain, immune cell infiltration and thickening of the synovial membrane in a dose-dependent manner. PWL recovered to near baseline level. Immunohistochemical analysis of spinal cord (CGRP, OX42) and dorsal root ganglia (TNFα) revealed dose-dependent reductions of pro-inflammatory biomarkers. Results showed 6.2 and 62 mg/day were effective doses. Exploratory behaviour was not altered by CBD indicating limited effect on higher brain function. CONCLUSIONS: These data indicate that topical CBD application has therapeutic potential for relief of arthritis pain-related behaviours and inflammation without evident side-effects.

Influence of supplemental, dietary vitamin A on retinol-binding protein concentrations in the plasma of preruminant calves.

Transport of retinol (vitamin A alcohol) from retinoid stores in the liver to target tissues is accomplished exclusively by a specific plasma protein, retinol-binding protein. Within individuals, retinol-binding protein concentrations in plasma are regulated and remain constant except in extremes of vitamin A nutriture or in disease. In the present study, retinol-binding protein concentrations in plasma from preruminant calves supplemented with 0, 1700 (i.e., current NRC requirement), 34,000, or 68,000 IU of vitamin A daily from birth to 27 d of age (n = 6/treatment) were quantified. Retinol-binding protein concentrations at birth averaged 21 microg/ml (n = 24) or approximately 50% of concentrations in dairy heifers and cows. Plasma retinol and retinol-binding protein concentrations were correlated positively, corroborating the role of vitamin A nutriture in the regulation of retinol-binding protein secretion from the liver. In this regard, dietary vitamin A influenced positively retinol and retinol-binding protein concentrations and, as a consequence, the degree of saturation of retinol-binding protein with retinol. At 27 d of age, calves fed > or = 34,000 IU of vitamin A had substantially higher retinol and retinol-binding protein concentrations than did calves fed < or = 1700 IU of vitamin A, indicating that dietary vitamin A effects positively vitamin A status. The data also suggest that the current NRC requirement may not be sufficient to assure vitamin A adequacy in preruminant calves. Percent saturation of retionol-binding protein with retinol in all calves was < 35%, much lower than anticipated and suggests that the retinol requirement of vitamin A-responsive tissues exceeded vitamin A availability.

Use of the relative dose response (RDR) assay to determine vitamin A status of calves at birth and four weeks of age.

An accurate assessment of vitamin A status can be determined by analysis of liver biopsy samples; however, liver biopsies are not always feasible. Plasma concentrations of vitamin A do not provide an accurate indication of vitamin A status. The objective of this study, therefore, was to determine the ability of the relative dose response assay to indicate the vitamin A status of Holstein calves. Calves were obtained at birth and assigned to vitamin A treatments (0, 1700, 34,000, or 68,000 IU/d) added to milk replacer. Liver biopsies and relative dose response assays were performed at birth and 4 wk. Calves supplemented with 1700, 34,000, or 68,000 IU of vitamin A/d had adequate (greater than 20 microg/g) liver concentrations of vitamin A at 4 wk of age. The relative dose response assay at 4 wk was correlated with liver concentrations of vitamin A. Both the relative dose response assay and liver concentrations of vitamin A indicated that calves not supplemented with vitamin A had low vitamin A status, whereas other treatment groups had adequate vitamin A status. Plasma concentrations of retinol increased by 4 wk of age in calves receiving supplemental vitamin A at 34,000 IU and 68,000 IU/d and decreased in unsupplemented calves; however, all calves had concentrations of <20 microg of retinol/dl of plasma. The relative dose response assay agreed with liver biopsies as an indication of vitamin A status, whereas plasma concentrations of retinol incorrectly indicated all treatment groups were deficient in vitamin A.

Effects of supplemental vitamin A on retinoic acid concentrations in the plasma of preruminant calves.

Neonatal calves are fed frequently milk replacers with vitamin A concentrations exceeding those recommended by the National Research Council. The vitamin A metabolite, retinoic acid (RA), affects profoundly cellular differentiation and homeostasis. For this reason, effects of dietary vitamin A on plasma concentrations of RA isomers in milk replacer-fed calves were examined. Male, Holstein calves (n = 24) were fed colostrum within 12 hours after birth and, thereafter, a custom-formulated low vitamin A milk replacer providing 0, 1700 [National Research Council (NRC) daily requirement for young growing calves] (controls), 34,000 (industry standard in the United States) or 68,000 IU of vitamin A daily. Concentrations of retinol and RA isomers in plasma samples collected from birth to 27 days of age were determined by HPLC. Retinol was affected by dietary vitamin A with higher concentrations occurring in calves supplemented with > or = 34,000 IU of vitamin A/day than in control (1700 IU of vitamin daily) and unsupplemented calves. Relative to controls, concentrations of all isomers of RA were higher in calves supplemented with > or = 34,000 of vitamin A daily during the experimental period. The predominant isomer in all calves was 9,13-dicis-RA. In control calves, 9,13-dicis-RA and 9-cis-RA were maximal at 1 to 6 days of age and then decreased progressively. In calves fed > or = 34,000 IU of vitamin A daily, concentrations of these isomers were markedly higher at 6 days of age, relative to controls, and remained elevated for the duration of the study. In all calves, retinol was correlated positively with 9,13-dicis- and 9-cis-RA from 9 to 27 days of age. 9,13-cis-Retinoic acid was correlated positively with 9-cis- and 13-cis-RA from 13 to 27 days of age. It is concluded that supplementing milk replacer-fed calves with vitamin A at levels exceeding current NRC recommendations by > or = 20-fold causes an elevation in plasma concentrations of retinol and retinoic acids. 9,13-dicis- and 9-cis-Retinoic acids were most affected by supplemental vitamin A. Physiologic consequences of increased plasma RA concentrations induced by high dietary levels of vitamin A warrant investigation.

Influence of vitamin A supplementation in milk on growth, health, concentrations of vitamins in plasma, and immune parameters of calves.

Seventy-seven Holstein calves were used to determine effects of vitamin A supplementation (0, 15,000, or 30,000 IU/d) of milk fed to calves through 6 wk of age. Effects of gender of calves and parity of dams also were considered. Supplementation with vitamin A did not affect retinol concentrations in plasma; however, calves fed milk containing supplemental vitamin A had decreased alpha-tocopherol concentrations in plasma at 6 wk compared with the concentrations in plasma of calves that were fed milk without supplemental vitamin A. Growth, serum protein, serum immunoglobulin (Ig) M and IgG, leukocyte proportions, and weekly fecal scores were not affected by vitamin A supplementation. Calves that scoured were fed milk supplemented with an additional 0 or 30,000 IU/d of vitamin A. Supplementation with an additional 30,000 IU/d when calves were scouring increased treatment days. Female calves had lower body measurements (weight, length, and height) at birth and greater fecal scores for wk 2 and 3 than did male calves. Gender did not affect serum protein, IgM, or IgG; however, female calves had higher percentages of monocytes and lower percentages of T cells than did male calves. At 6 wk, female calves also had higher percentages of B cells than did male calves. These data indicate that ratios of vitamins A and E should be considered in dietary formulations for calves. Also, additional vitamin A provided by some scour treatments could be detrimental to calves that are already receive vitamin A supplementation.

Effect of vitamins A and E on nitric oxide production by blood mononuclear leukocytes from neonatal calves fed milk replacer.

This study evaluated the effects of dietary vitamin A and E on the in vitro capacity of blood mononuclear leukocytes from calves to produce nitric oxide. Calves fed milk replacer received 100 IU/d of vitamin E as RRR-alpha-tocopherol or RRR-alpha-tocopheryl acetate and 0, 1700, 34,000, or 68,000 IU of vitamin A as retinyl acetate. Leukocytes from calves produced greater amounts of nitric oxide relative to leukocytes from adult cattle. The greater production of nitric oxide by calf leukocytes may be typical of the immature neonatal immune system. Nitric oxide production by calves fed RRR-alpha-tocopherol and either 1700 or 34,000 IU of vitamin A was less than that of calves in other groups and was more typical of production by leukocytes from cows. Our data suggest that optimal amounts of dietary vitamins A and E prompt the maturation of this response toward one that is more typical of adult cattle. Leukocytes from 1-wk-old calves produced less nitric oxide and were less responsive to stimuli than were leukocytes from older calves, a possible consequence of suppressive factors that were present in the ingested colostrum or in the circulation at birth.