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Combination of platelet-rich plasma and bone marrow mesenchymal stem cells enhances tendon – bone healing in a rabbit model of anterior cruciate ligament reconstruction

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ABSTRACT

Background: The objective of this study was to investigate the potency of platelet-rich plasma (PRP) combined with bone marrow mesenchymal stem cells (BMSCs) to promote tendon–bone healing in a rabbit model.

Methods: In the in vitro study, the effects of PRP on osteogenic induction of BMSCs were analysed. Later, PRP with or without BMSCs was used in the rabbit model of anterior cruciate ligament reconstruction. Specimens were harvested 8 weeks postoperatively to evaluate tendon–bone healing by histology, radiology, and biomechanical testing.

Results: The in vitro study revealed that collagen I, osteocalcin, and osteopontin expression was higher in BMSCs co-cultured with PRP for 14 days. The in vivo study revealed a more mature tendon–bone interface using light microscopy, a more newly formed bone at the bone tunnel walls detected by micro-computed tomography, and a significantly higher failure load as assessed by biomechanical testing in the BMSC + PRP group than in the control and PRP groups.

Conclusions: These results indicate that the combination of PRP and BMSCs promotes tendon–bone healing and has potential for clinical use.

Electronic supplementary material: The online version of this article (doi:10.1186/s13018-016-0433-7) contains supplementary material, which is available to authorized users.

No MeSH data available.


a Gross observations of the semitendinosus tendon (black arrow) and b native anterior cruciate ligament (ACL) (black arrow). c Macroscopic view of ACL reconstruction (arrow points to autologous semitendinosus tendon graft)
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Fig1: a Gross observations of the semitendinosus tendon (black arrow) and b native anterior cruciate ligament (ACL) (black arrow). c Macroscopic view of ACL reconstruction (arrow points to autologous semitendinosus tendon graft)

Mentions: Animal experiments were approved by the Zhejiang University Ethics Committee. A total of 30 New Zealand White rabbits (age, 12 weeks, and weight, 2.5 ± 0.2 kg) were used in this study. All rabbits underwent ACL reconstruction in the left hind leg after intravenous anaesthesia with 30 mg/kg body weight pentobarbital sodium solution (Dawen Biotech, Seoul, Korea). After shaving and disinfecting the left hind leg, a lateral parapatellar incision was made to expose the knee joint. After the native ACL was excised, the tibial and femoral bone tunnels were created with a 2.5-mm-diameter drill (Fig. 1b). The ipsilateral semitendinosus tendon was harvested to reconstruct the ACL (Fig. 1a). The rabbits were divided randomly into three groups. In the first group, a normal hamstring tendon was used for ACL reconstruction (control group). Rabbits from the second group (PRP group) received hamstring tendons wrapped with 0.1 mL PRP immobilised in 0.1 mL fibrin glue (TISSEEL kit; Baxter AG, Vienna, Austria) for ACL reconstruction. In the third group (BMSC + PRP group), 1 × 107 BMSCs were immobilised in 0.2 mL PRP and 0.1 mL fibrin glue and used for ACL reconstruction. The PRP concentration and number of BMSCs were determined by flow cytometry. Grafts for the PRP and BMSC + PRP groups were wrapped with glue immediately before insertion into the bone tunnels. Both ends of the graft were fixed by sutures tied over screws in the femur and tibia (Fig. 1c). Animals were allowed to move freely postoperatively. All rabbits were sacrificed with a lethal injection of pentobarbital 8 weeks postoperatively for assessment; half of the specimens in each group (n = 5/group) were used for the histological assessment and the other half (n = 5/group) were used for radiological and biomechanical assessments.Fig. 1


Combination of platelet-rich plasma and bone marrow mesenchymal stem cells enhances tendon – bone healing in a rabbit model of anterior cruciate ligament reconstruction
a Gross observations of the semitendinosus tendon (black arrow) and b native anterior cruciate ligament (ACL) (black arrow). c Macroscopic view of ACL reconstruction (arrow points to autologous semitendinosus tendon graft)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5015347&req=5

Fig1: a Gross observations of the semitendinosus tendon (black arrow) and b native anterior cruciate ligament (ACL) (black arrow). c Macroscopic view of ACL reconstruction (arrow points to autologous semitendinosus tendon graft)
Mentions: Animal experiments were approved by the Zhejiang University Ethics Committee. A total of 30 New Zealand White rabbits (age, 12 weeks, and weight, 2.5 ± 0.2 kg) were used in this study. All rabbits underwent ACL reconstruction in the left hind leg after intravenous anaesthesia with 30 mg/kg body weight pentobarbital sodium solution (Dawen Biotech, Seoul, Korea). After shaving and disinfecting the left hind leg, a lateral parapatellar incision was made to expose the knee joint. After the native ACL was excised, the tibial and femoral bone tunnels were created with a 2.5-mm-diameter drill (Fig. 1b). The ipsilateral semitendinosus tendon was harvested to reconstruct the ACL (Fig. 1a). The rabbits were divided randomly into three groups. In the first group, a normal hamstring tendon was used for ACL reconstruction (control group). Rabbits from the second group (PRP group) received hamstring tendons wrapped with 0.1 mL PRP immobilised in 0.1 mL fibrin glue (TISSEEL kit; Baxter AG, Vienna, Austria) for ACL reconstruction. In the third group (BMSC + PRP group), 1 × 107 BMSCs were immobilised in 0.2 mL PRP and 0.1 mL fibrin glue and used for ACL reconstruction. The PRP concentration and number of BMSCs were determined by flow cytometry. Grafts for the PRP and BMSC + PRP groups were wrapped with glue immediately before insertion into the bone tunnels. Both ends of the graft were fixed by sutures tied over screws in the femur and tibia (Fig. 1c). Animals were allowed to move freely postoperatively. All rabbits were sacrificed with a lethal injection of pentobarbital 8 weeks postoperatively for assessment; half of the specimens in each group (n = 5/group) were used for the histological assessment and the other half (n = 5/group) were used for radiological and biomechanical assessments.Fig. 1

View Article: PubMed Central - PubMed

ABSTRACT

Background: The objective of this study was to investigate the potency of platelet-rich plasma (PRP) combined with bone marrow mesenchymal stem cells (BMSCs) to promote tendon–bone healing in a rabbit model.

Methods: In the in vitro study, the effects of PRP on osteogenic induction of BMSCs were analysed. Later, PRP with or without BMSCs was used in the rabbit model of anterior cruciate ligament reconstruction. Specimens were harvested 8 weeks postoperatively to evaluate tendon–bone healing by histology, radiology, and biomechanical testing.

Results: The in vitro study revealed that collagen I, osteocalcin, and osteopontin expression was higher in BMSCs co-cultured with PRP for 14 days. The in vivo study revealed a more mature tendon–bone interface using light microscopy, a more newly formed bone at the bone tunnel walls detected by micro-computed tomography, and a significantly higher failure load as assessed by biomechanical testing in the BMSC + PRP group than in the control and PRP groups.

Conclusions: These results indicate that the combination of PRP and BMSCs promotes tendon–bone healing and has potential for clinical use.

Electronic supplementary material: The online version of this article (doi:10.1186/s13018-016-0433-7) contains supplementary material, which is available to authorized users.

No MeSH data available.