Abstract
Context Sanguisorba officinalis L. (Rosaceae), a famous traditional Chinese medicine. It was recently reported that its polysaccharide could facilitate collagen production. Objectives We investigated the mechanism by which S. officinalis polysaccharide (SOWPa) and/or platelet-rich plasma (PRP) promote regenerative potential of anterior cruciate ligament (ACL) in vitro. Materials and methods ACL fibroblasts were treated with SOWPa (25 and 100 mg/kg), PRP, PRP + SOWPa (25 and 100 mg/kg) or vehicle alone for 24, 48, or 72 h. Cell viability, migration ability and apoptosis were evaluated by MTT, transwell and flow cytometry, respectively. Western blot analysis was performed to assess associated protein expression. Results PRP, SOWPa (100 mg/kg) or PRP + SOWPa (100 mg/kg) treatment for 72 h significantly improved the cell viability of ACL fibroblasts from 100 ± 7.5% (control) to 156.85 ± 12.82%, 188.08 ± 15.92%, and 223.67 ± 18.82%, respectively, which was evidenced by individual decreased apoptosis rate from 31.26 ± 2.35% (control) to 20.80 ± 1.89%, 18.01 ± 1.55% and 9.33 ± 0.78%. Furthermore, the motility of ACL fibroblasts was significantly improved with increased migrated cell number per field from 5 for control to 26 for PRP, 36 for SOWPa and 44 for PRP + SOWPa, respectively. Moreover, the protein expression of differentiation markers (RUNX2, ALP, BMP2 and Col I) and TLR-4 and phosphorylated p65 (p-p65) was inhibited by the above treatment. Discussion and conclusions Data suggested that the addition of SOWPa to PRP increased the regenerative ability of ACL fibroblasts by blocking the TLR-4/NF-κB pathway.
Highlights
The anterior cruciate ligament (ACL) is an important intraarticular structure to maintain the stability of knee joint and its rupture causes pain and knee instability, and runs the risk of premature degenerative joint disease (Liden et al 2008)
A water-soluble polysaccharide SOWPa was purified from the roots of S. officinalis, by lipid removal with 80% ethanol, hot treated with platelet-rich plasma (PRP) þ SOWPa than either SOWPa or PRP treatment at any time
To further confirm and quantify apoptosis ACL fibroblasts after treatment with PRP, SOWPa (100 mg/kg) or PRP þ SOWPa (100 mg/kg), the cells were stained with Annexin V-FITC/PI and analysed by flow cytometry
Summary
The anterior cruciate ligament (ACL) is an important intraarticular structure to maintain the stability of knee joint and its rupture causes pain and knee instability, and runs the risk of premature degenerative joint disease (Liden et al 2008). Owing to the shortage of soft-tissue autografts/allografts tissue, various synthetic, nondegradable allografts or autografts were used in the 1970s and 1980s but were hampered by their inherent limitations such as premature graft rupture, foreign body reactions, osteolysis and synovitis (De Groot et al 1996) To overcome these aforementioned problems, there is an increasing interest in strategies to improve the healing of ACL with minimal surgical interference, such as the use of platelet-rich plasma (PRP)
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