Human amniotic suspension allograft positively affect tendon healing in a rat model of collagenase-induced tendinopathy
Author(s):
de Girolamo Laura (Italy)
de Girolamo Laura (Italy)
Affiliations:
ESSKA Academy. DE GIROLAMO L. 05/09/18; 209854; P20-1529 Topic: Basic Science
Dr. Laura DE GIROLAMO
Dr. Laura DE GIROLAMO
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Abstract
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Objectives: Different growth factors and cytokines, such as insulin-like growth factors (IGFs) and platelet-derived growth factors (PDGFs) are known to be involved in normal tendon healing. In this view, placental derived tissues might be useful in promoting tendon regeneration as they contain a number of teno-conductive growth factors and cytokines, including IGFs, PDGFs, and other regenerative and anti-inflammatory factors. In particular, human amniotic suspension allograft (ASA), containing amniotic membrane particulate and cells from the amniotic fluid, has shown promising results in orthopaedic regenerative medicine. The aim of this study was to evaluate the efficacy of ASA in an already established Achilles tendinopathy model; the hypothesis is that the use of ASA would improve tendon healing compared to untreated controls.

Methods: A collagenase solution was injected along the right Achilles tendon of 78 Sprague Dawley rats; the contralateral limb was either left untreated or injected with saline solution. Seven days later, the right Achilles tendons were either left untreated (control) or treated with saline, or ASA injection. The rats were then sacrificed 14 and 28 days post-treatment and tendons were assessed for histology (Hematoxylin and Eosin (H&E) staining) and scored. Additionally at 28 days biomechanical testing was conducted on retrieved tendons to measure tensile strength. Immunohistochemistry was also conducted to detect the presence of collagen III and human nuclei within retrieved tendons.

Results: As showed by histological scoring, a single injection of ASA was able to improve significantly the tendon healing of collagenase-treated Achilles tendons. In particular, an increased fiber alignment and decreased cell infiltration were observed in ASA-treated animals compared to untreated or saline-treated ones. However, despite histological differences, biomechanical testing did not reveal significant improvements between tendons from rats treated with or without ASA. The presence of Collagen III was slightly downregulated in the tendons of rats who received ASA compared to controls. Interestingly, after 28 days from the treatment, anti-human nuclei positive staining suggested the presence of components from ASA in the tendon.

Conclusions: The injured Achilles tendons treated with ASA showed structural improvement in comparison to controls. Although there were no differences in the biomechanical maximum load between tendons treated with or without ASA, collagen III staining was slightly downregulated in ASA-treated rats. These findings suggest a concrete role of ASA in promoting a more efficient tendon healing in this model of Achilles tendinopathy in rats. Although this data supports the regenerative and anti-inflammatory properties of amniotic tissues, additional clinical work evaluating ASA for tendinopathy is required.

Keywords:
Amniotic suspension allograft; achilles tendinopathy; tendon; regenerative medicine
Objectives: Different growth factors and cytokines, such as insulin-like growth factors (IGFs) and platelet-derived growth factors (PDGFs) are known to be involved in normal tendon healing. In this view, placental derived tissues might be useful in promoting tendon regeneration as they contain a number of teno-conductive growth factors and cytokines, including IGFs, PDGFs, and other regenerative and anti-inflammatory factors. In particular, human amniotic suspension allograft (ASA), containing amniotic membrane particulate and cells from the amniotic fluid, has shown promising results in orthopaedic regenerative medicine. The aim of this study was to evaluate the efficacy of ASA in an already established Achilles tendinopathy model; the hypothesis is that the use of ASA would improve tendon healing compared to untreated controls.

Methods: A collagenase solution was injected along the right Achilles tendon of 78 Sprague Dawley rats; the contralateral limb was either left untreated or injected with saline solution. Seven days later, the right Achilles tendons were either left untreated (control) or treated with saline, or ASA injection. The rats were then sacrificed 14 and 28 days post-treatment and tendons were assessed for histology (Hematoxylin and Eosin (H&E) staining) and scored. Additionally at 28 days biomechanical testing was conducted on retrieved tendons to measure tensile strength. Immunohistochemistry was also conducted to detect the presence of collagen III and human nuclei within retrieved tendons.

Results: As showed by histological scoring, a single injection of ASA was able to improve significantly the tendon healing of collagenase-treated Achilles tendons. In particular, an increased fiber alignment and decreased cell infiltration were observed in ASA-treated animals compared to untreated or saline-treated ones. However, despite histological differences, biomechanical testing did not reveal significant improvements between tendons from rats treated with or without ASA. The presence of Collagen III was slightly downregulated in the tendons of rats who received ASA compared to controls. Interestingly, after 28 days from the treatment, anti-human nuclei positive staining suggested the presence of components from ASA in the tendon.

Conclusions: The injured Achilles tendons treated with ASA showed structural improvement in comparison to controls. Although there were no differences in the biomechanical maximum load between tendons treated with or without ASA, collagen III staining was slightly downregulated in ASA-treated rats. These findings suggest a concrete role of ASA in promoting a more efficient tendon healing in this model of Achilles tendinopathy in rats. Although this data supports the regenerative and anti-inflammatory properties of amniotic tissues, additional clinical work evaluating ASA for tendinopathy is required.

Keywords:
Amniotic suspension allograft; achilles tendinopathy; tendon; regenerative medicine
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