Application of Exogenous Esterified Hyaluronan to Equine Distal Limb Wounds

Abstract 

The objective of this study was to evaluate the efficacy of topical application of a hyaluronan (HA) derivative in wound healing with respect to the rate of epithelialization, fibroplasia, angiogenesis and contraction, magnitude of the local inflammatory response, local expression of transforming growth factor-β 1 and 3 (TGF-β 1 and 3), tumor necrosis factor-α (TNF-α), and collagen type III deposition. In six healthy adult horses, six full-thickness skin wounds were created on the dorsal aspect of both metacarpi using a sterile template. Sites were sampled at 0, 1, 2, 5, 14, 21, and 35 days following wounding. Wounds on one limb were dressed with commercially available esterified HA fleece under a nonadherent dressing. The opposite limb was covered with the nonadherent dressing alone (control). Images of the most proximal wounds were used to determine the area of total healing and the relative contributions of epithelialization and contraction to healing. At each sample time, a control and treatment biopsy were taken for histological evaluation and special stains. All samples were evaluated for degree of inflammation, fibroplasia and angiogenesis; in situ hybridization for type III collagen, TGFβ1 and 3, and immunohistochemistry for TNF-α. Mean percentages of total wound healing, epithelialization, and wound contraction were not significantly different between control and treatment groups. In treated horses, initial wound expansion was significantly decreased during the first 2 weeks. Mononuclear cell numbers, counted in the granulation tissue, increased in both control and treated limbs over the entire course of the study. However at day 35 the macrophage numbers counted in the treated horses were significantly increased compared with the control limbs (P < .05). Although not statistically significant, relative staining for type III collagen in the treated wounds was less than that of control wounds. Results of the present study do not support a benefit of an exogenous HA-derivative in the healing of distal limb wounds in horses. The shortcomings of the study design are discussed.

Keywords: Equine, Wound healing, Hyaluronan, Lower limb