Surfaces Covered with Polylactic Acid Nanoparticles: as a Safe Method for the Production of Blood Derivatives
Surfaces Covered with Polylactic Acid Nanoparticles: as a Safe Method for the Production of Blood Derivatives
Majid Zamani,1Saeid Kaviani,2,*Mehdi Yousefi,3Saeid Abroun,4Mohammad Hojjat-Farsangi,5Behzad Pourabbas,6
1. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 3. Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 4. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 5. Bioclinicum, Department of Oncology-Pathology, Karolinska Institute, Bioclinicum, Stockholm, Sweden 6. Department of Polymer Engineering, Sahand University of Technology, Tabriz, Iran
Introduction: In recent years, the use of blood derivatives in regenerative medicine has attracted the attention of researchers. Various blood derivatives such as platelet-rich plasma (PRP), autologous conditioned serum (ACS), platelet lysate (PL), and platelet-rich fibrin (PRF) are used in tissue regeneration. In these blood derivatives, blood cells and secreted growth factors from these cells are used for tissue regeneration in wound healing, bone fracture, osteoarthritis, and tendon injuries. In different blood derivatives, different methods are used to activate cells and release the growth factors of these cells. Compounds such as thrombin, calcium chloride (CaCl2), collagen and even glass beads can be used to activate blood cells and release growth factors from these cells. In ACS, glass beads contact with blood cells, including monocytes, causing the activation of these cells, and various cytokines, especially anti-inflammatory cytokines, are released from these cells. Polylactic acid (PLA) is one of the compounds approved by the U.S. Food and Drug Administration (FDA), which has been used in various medical fields, and one of its side effects is the activation of various blood cells, including platelets and monocytes. The effect of PLA on blood cells can be used to activate these cells and release growth factors in blood derivatives. By coating glass beads with PLA, it is possible to provide a suitable surface for the activation of blood cells to increase the amount of growth factors and cytokines of the blood derivatives and prevent the release of PLA into the sample and prevent its negative effects for patients. In this study, we intend to coat the borosilicate glass beads with PLA nanoparticles and investigate the coating of the beads, the non-release of PLA nanoparticles inside the sample, and the hemolysis of the sample, so that it can be used for the production of blood products for regenerative medicine purposes in the future.
Methods: PLA nanoparticles were used to coat medical-grade borosilicate beads with a diameter of 3 ml, and the coating of glass beads was evaluated by scanning electron microscopy (SEM). To investigate the effect of coated beads, after obtaining informed consent, 17 ml of blood was taken in two tubes (8.5 ml in each tube, one of the tubes had beads covered with PLA nanoparticles and the other tube had no willows to prepare PRP) containing 3 ml of anticoagulant citrate dextrose-A (ACD-A) (1.5 ml anticoagulant in each tube) from 7 male volunteers who meet the entry criteria including: age 20-50 years, platelet count 150-400×103/ml, leukocyte count 6-10×103/ml, no underlying diseases, did not consume alcohol, drugs, anticoagulant and immunosuppressive drugs, no history of chemotherapy, and smoking. The tube without the coated beads was immediately centrifuged at 150 g for 10 minutes and the supernatant, which was PRP, was separated. The tube containing coated beads was incubated for 6 hours at 37 C°, then the samples were centrifuged at 150g for 10 minutes and the upper layer containing the activated plasma was separated. Fourier-transform infrared spectroscopy (FT-IR) was used to evaluate the release of PLA nanoparticles into the activated plasma and hemoglobin measurement was used to evaluate the hemolysis of the sample.
Results: SEM images showed that the surface of the beads was properly coated with PLA nanoparticles. FT-IR was used to evaluate the release of PLA nanoparticles into the conditioned plasma, which showed that these particles were not released into the sample. Hemoglobin measurement showed that coated beads did not cause hemolysis and the amount of activated plasma hemoglobin did not increase compared to PRP.
Conclusion: PLA-coated glass beads can be used to prepare blood derivatives without releasing them into the conditioned plasma and hemolysis of the sample. This study investigated the negative effects of these coated beads on the blood derivatives, and investigating the effectiveness of these coated beads in activating cells and increasing the amount of growth factors and cytokines in the activated plasma requires further studies and evaluation of its effectiveness on tissue regeneration.