Changes in biological characteristics of platelet-rich fibrin by freeze-drying technology

doi:10.12307/2021.142

Abstract

Abstract: BACKGROUND: Platelet-rich fibrin has been used in bone and soft tissue repair in clinic. However, due to its biological activity, it is limited to short-term clinical application. Therefore, how to keep platelet-rich fibrin stably and prolong the time of clinical application is very important.
OBJECTIVE: To explore the effect of freeze-drying technology on the morphology of platelet-rich fibrin and the activity and release regularity of growth factor releasing from platelets in platelet-rich fibrin.
METHODS: Fresh and freeze-dried platelet-rich fibrin were prepared and then soaked in DMEM basic medium without fetal bovine serum. The supernatant of freeze-dried or fresh platelet-rich fibrin was collected at 1, 7, 14 and 21 days and used for fibroblast culture and platelet-derived growth factor-BB content determination. The morphological structure, activity and release regularity of growth factor releasing from platelets in platelet-rich fibrin were compared before and after freeze-drying. Morphological observation was detected by hematoxylin-eosin staining. Cell proliferation was measured by CCK8 assay. The cell migration ability was evaluated by cell scratch test. Platelet-derived growth factor-BB content of collected supernatant at different time points in the two groups was detected by using ELISA kit.
RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining showed that the structure of freeze-dried platelet-rich fibrin was loose and spongy, and the pore diameter was large. The number of nuclear cells in blue staining was significantly reduced. (2) Compared with the serum-free medium, the fresh and freeze-dried platelet-rich fibrin extracts collected on the 1 and 7 days significantly promoted the proliferation of fibroblasts (P < 0.05), and freeze-dried platelet-rich fibrin extracts strongly promoted the proliferation capacity of fibroblasts (P < 0.05). (3) Compared with the serum-free medium, the fresh and freeze-dried platelet-rich fibrin extract, and the complete medium containing 10% fetal bovine serum could effectively promote the migration of fibroblasts at 12 and 24 hours, and the difference was statistically significant (P < 0.05). (4) Compared with the fresh platelet-rich fibrin, the freeze-dried platelet-rich fibrin showed higher migration ability of fibroblasts at 12 and 24 hours (P < 0.05), and the difference was statistically significant (P < 0.05). (5) The content of platelet-derived growth factor-BB in fresh platelet-rich fibrin group was the largest at 7 days, and the release of growth factor gradually decreased. The content of platelet-derived growth factor-BB in freeze-dried platelet-rich fibrin group was the largest at 1 day, and the release of growth factor gradually decreased with time, and the difference was statistically significant (P < 0.05). The total content of platelet-derived growth factor-BB showed no significant difference between the two groups (P > 0.05). (6) The results show that freeze-drying technology is feasible for platelet-rich fibrin preservation, which can provide theoretical basis for tissue repair and reconstruction.

Key words: freeze-drying technology, platelet-rich fibrin, fibroblasts, platelet-derived growth factor BB, cell proliferation, cell migration

CLC Number:

Liu Lei, Di Haiping, Guo Haina, Cao Dayong, Niu Xihua, Xia Chengde. Changes in biological characteristics of platelet-rich fibrin by freeze-drying technology[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(31): 4995-4999.

Figures/Tables 4

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