Macroscopic, Histologic, and Immunomodulatory Response of Limb Wounds Following Intravenous Allogeneic Cord Blood-Derived Multipotent Mesenchymal Stromal Cell Therapy in Horses.

Limb wounds are common in horses and often develop complications. Intravenous multipotent mesenchymal stromal cell (MSC) therapy is promising but has risks associated with intravenous administration and unknown potential to improve cutaneous wound healing. The objectives were to determine the clinical safety of administering large numbers of allogeneic cord blood-derived MSCs intravenously, and if therapy causes clinically adverse reactions, accelerates wound closure, improves histologic healing, and alters mRNA expression of common wound cytokines. Wounds were created on the metacarpus of 12 horses. Treatment horses were administered 1.51-2.46 × 108 cells suspended in 50% HypoThermosol FRS, and control horses were administered 50% HypoThermosol FRS alone. Epithelialization, contraction, and wound closure rates were determined using planimetric analysis. Wounds were biopsied and evaluated for histologic healing characteristics and cytokine mRNA expression. Days until wound closure was also determined. The results indicate that 3/6 of treatment horses and 1/6 of control horses experienced minor transient reactions. Treatment did not accelerate wound closure or improve histologic healing. Treatment decreased wound size and decreased all measured cytokines except transforming growth factor-ß3. MSC intravenous therapy has the potential to decrease limb wound size; however, further work is needed to understand the clinical relevance of adverse reactions.

Homing and Engraftment of Intravenously Administered Equine Cord Blood-Derived Multipotent Mesenchymal Stromal Cells to Surgically Created Cutaneous Wound in Horses: A Pilot Project.

Limb wounds on horses are often slow to heal and are prone to developing exuberant granulation tissue (EGT) and close primarily through epithelialization, which results in a cosmetically inferior and non-durable repair. In contrast, wounds on the body heal rapidly and primarily through contraction and rarely develop EGT. Intravenous (IV) multipotent mesenchymal stromal cells (MSCs) are promising. They home and engraft to cutaneous wounds and promote healing in laboratory animals, but this has not been demonstrated in horses. Furthermore, the clinical safety of administering >1.00 × 108 allogeneic MSCs IV to a horse has not been determined. A proof-of-principle pilot project was performed with two horses that were administered 1.02 × 108 fluorescently labeled allogeneic cord blood-derived MSCs (CB-MSCs) following wound creation on the forelimb and thorax. Wounds and contralateral non-wounded skin were sequentially biopsied on days 0, 1, 2, 7, 14, and 33 and evaluated with confocal microscopy to determine presence of homing and engraftment. Results confirmed preferential homing and engraftment to wounds with persistence of CB-MSCs at 33 days following wound creation, without clinically adverse reactions to the infusion. The absence of overt adverse reactions allows further studies to determine effects of IV CB-MSCs on equine wound healing.

Sinocutaneous fistula repair with a masseter muscle transposition flap combined with wound matrix and cancellous bone graft in a horse: A new technique.

OBJECTIVE: To describe a new technique to repair a sinocutaneous fistula with a masseter muscle transposition flap. STUDY DESIGN: Case report. ANIMAL: One 13-year-old thoroughbred stallion. METHODS: One 13-year-old stallion with a 3.5 × 6-cm sinocutaneous fistula over the right caudal maxillary sinus was treated with a transpositional masseter muscle flap. This repair consisted of a commercial wound matrix dressing placed directly over the hole in the maxilla and secured with suture material; a cancellous bone graft collected from the right tuber coxa placed on the dressing; and a portion of the superficial layer of the masseter muscle, with its pedicle at the facial crest, transposed dorsally over the bone graft, followed by a rotational skin flap with skin rostral to the fistula to close the defect. RESULTS: Seroma formation and dehiscence of the skin flap occurred, but the transposed muscle flap survived, and the technique resulted in successful closure of the sinocutaneous fistula with excellent cosmetic and functional outcome. CONCLUSION: A chronic maxillary sinocutaneous fistula was successfully treated by using a transposition flap of the masseter muscle and a rotational skin flap with minor complications. CLINICAL IMPACT: Transposition of the superficial layer of the masseter muscle should be considered for a repair of large maxillary sinocutaneous fistulas in horses.

Identification of mRNA of the Inflammation-associated Proteins CXCL8, CXCR2, CXCL10, CXCR3, and ß-Arrestin-2 in Equine Wounded Cutaneous Tissue: a Preliminary Study.

Horses often sustain cutaneous wounds and healing can be prolonged and difficult to treat. Compared to body wounds, limb wounds heal slower and are more likely to develop exuberant granulation tissue. Differences in healing rates and exuberant granulation tissue formation is attributed to abnormal cytokine profiles. CXCL8 and its receptor CXCR2 are involved in acute inflammation whereas CXCL10 and its receptor CXCR3 are involved in inflammation resolution. ß- arrestin-2 regulates inflammation through internalization of G-protein coupled receptors (GPCRs) including CXCR2 and CXCR3. Gene expression of these five inflammation associated proteins have not been previously identified in equine cutaneous tissue and may play a role in dysregulation of inflammation in equine limb wounds. The mRNA expression levels were measured using QuantiGene Plex Assay from cutaneous biopsies collected from surgically created wounds on the limb and thorax on days 0, 1, 2, 7, 14, and 33 from two horses. The mRNA expression levels were measured in mean fluorescent intensity and graphed. We were successful in identifying all five proteins for the first time in equine cutaneous tissue. Preliminary results suggest that there are different expression patterns for CXCL8, CXCR2 and ß-arrestin-2 between the limb and thorax but not for CXCL10 and CXCR3. Differential regulation of CXCL8, CXCR2 and ß-arrestin-2 may further explain why limb wounds heal differently than body wounds and warrants further investigation.