Abstract
BackgroundThis pilot study aimed to ascertain whether the local application of ascorbic acid (AA), of T3, and of rat (r) bone marrow mesenchymal stem cells (BMSCs), alone or in all possible combinations, promoted healing after an Achilles tendon injury in a rat model.MethodsAn Achilles tendon defect was produced in 24 6–8-week-old male inbred Lewis rats. The animals were then randomly divided into eight groups of three rats each. The tendon defect was filled with 50 μL of phosphate-buffered saline (PBS) containing (1) 50 μg/mL AA (AA group), (2) 10−7 M T3 (T3 group), (3) 4 × 106 rBMSCs (rBMSC group), (4) 50 μg/mL AA + 10−7 M T3 (AA + T3 group), (5) 4 × 106 rBMSCs + 50 μg/mL AA (rBMSC + AA group), (6) 4 × 106 rBMSCs + 10−7 M T3 (rBMSC + T3 group), (7) 4 × 106 rBMSCS + 50 μg/mL AA + 10−7 M T3 (rBMSC + AA + T3 group), and (8) PBS only (control group: CTRL). All treatments were administered by local injection immediately after the tendons had been damaged; additionally, AA was injected also on the second and fourth day from the first injection (for groups 1, 4, 5, and 7), and T3 was injected again every day for 4 days (for groups 2, 4, 6, and 7). At 30 days from initial treatment, tendon samples were harvested, and the quality of tendon repair was evaluated using histological and histomorphological analysis. The structure and morphology of the injured Achilles tendons were evaluated using the modified Svensson, Soslowsky, and Cook score, and the collagen type I and III ratio was calculated.ResultsThe group treated with AA combined with T3 displayed the lowest Svensson, Soslowsky, and Cook total score value of all tissue sections at histopathological examination, with fiber structure close to regular orientation, normal-like tendon vasculature, and no cartilage formation. AA + T3 also showed the highest collagen I and the lowest collagen III values compared to all other treatments including the CTRL.ConclusionThere are potential benefits using a combination of AA and T3 to accelerate tendon healing.
Highlights
About 3 to 5 million patients worldwide experience tendon injuries each year [1]
Identification of rBMSC Once they had been isolated and cultured, the rBMSCs were analyzed for immunophenotype by flow cytometry, which confirmed the expression of CD90 and the lack of CD45 (Fig. 1a)
The results indicate that ascorbic acid acts in synergy with T3 to accelerate tendon healing
Summary
About 3 to 5 million patients worldwide experience tendon injuries each year [1]. Tendon healing is slow and often incomplete: the currently available techniques for surgical intervention and repair are inadequate [2,3,4,5]. BMSCs alone seem ineffective [9, 10], and recent biological approaches, such as BMSCs in combination with growth factors, and specialized delivery systems are being explored to enhance tendon healing [11,12,13]. This pilot study aimed to ascertain whether the local application of ascorbic acid (AA), of T3, and of rat (r) bone marrow mesenchymal stem cells (BMSCs), alone or in all possible combinations, promoted healing after an Achilles tendon injury in a rat model
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