Platelet Rich Plasma From Umbilical Cord Blood With Chinese Medicine Formula Ling Zhi And San-miao-san In Hyaluronic Gel For Articular Cartilage Retrieval In Rat Osteoarthritis Model

Abstract

Background: Osteoarthritis (OA) is the most common degenerative joint disease. Umbilical cord blood (UCB) Platelet Rich Plasma (PRP) is an autologous, non-immunogenic concentrate of platelets and a rich source of growth factors. Hyaluronic acid (HA) gel scaffolds function as a delivery vehicle, which augment platelet viability in UCB PRP with the traditional Chinese medicine formulas Ling zhi (LZ) and San-Miao-San (SMS). Our aim is to address this issue and UCB PRP therapy, specifically using the potential therapeutic activities of LZ-SMS, which has been used empirically for the treatment of OA, to promote chondrocyte proliferation and exhibit additive or synergistic immunoregulatory effects for the articular cartilage retrieval at a rodent model of traumatic OA and subsequent cartilage damage. Subjects and Methods: Selected patients undergoing total knee arthroplasty (TKA) for OA and equal age- and sex-matched healthy controls were recruited and patients' demographic data of age, drugs and Oxford knee scores were recorded. The radiographs of OA knee were evaluated according to the Kellegren and Lawrence system. The biochemical parameters from blood and synovial fluid samples were measured. The validity of the histopathology grading systems was also examined using the Société Française d'Arthroscopie chondropathy scoring system. Synovial membrane and articular cartilage were obtained from patients undergoing TKA. In the vivo study, the traumatic OA animal model was made by transection of the anterior cruciate ligament (ACLT) in Sprague-Dawley rats and resulting in a true instability-induced OA lesion that mimics human traumatic OA. All ACLT treated rats were administrated with UCB PRP, UCB PRP HA gel, and UCB PRP with LZ-SMS HA gel, respectively, twice a week for 3 consecutive months, following which the animals were performed micro-CT scan, biochemical parameters detection and histological staining. Results: We obtained an average PRP concentration of around 1 × 107 platelets/ml. The site of injection was first confirmed using a rat cadaver and verifying with dye. We tagged platelets with the cell tracker indium-111 radioisotope and then used a confocal microscope to view the platelet distributed throughout in the HA gel. We found that human endothelial cells cultured with thrombin activated PRP induced relatively more endothelial cell proliferation compared to control, which demonstrated the potential benefit of PRP to induce cartilage proliferation. In our pilot vivo study, the female Dukin Hartley guinea pigs treated with 100 μl of PRP HA gel were not found any significant worsening in morphology of cartilage, when compared to the control limb. Furthermore, the treated limb had an increased content of glycosaminoglycan compared to the control limb. Discussion and Conclusion: The study is to elucidate the in vivo immunoregulatory effect of the refined UCB PRP with LZ-SMS on the articular cartilage retrieval in a rodent model of traumatic OA and promotes chondrocyte proliferation in vitro, as well as characterizing the clinical features of OA patients. We now wish to investigate further the effect of UCB PRP with LZ-SMS in HA gels on OA cartilage retrieval and chondrocytes proliferation.