Abstract
In 2004, we developed autologous periosteal sheets for the treatment of periodontal bone defects. This regenerative therapy has successfully regenerated periodontal bone and augmented alveolar ridge for implant placement. However, the necessity for 6-week culture is a limitation. Here, we examined the applicability of a human platelet-rich fibrin extract (PRFext) as an alternative to fetal bovine serum (FBS) for the explant culture of periosteal sheets in a novel culture medium (MSC-PCM) originally developed for maintaining mesenchymal stem cells. Small periosteum tissue segments were expanded in MSC-PCM + 2% PRFext for 4 weeks, and the resulting periosteal sheets were compared with those prepared by the conventional method using Medium199 + 10% FBS for their growth rate, cell multilayer formation, alkaline phosphatase (ALP) activity, and surface antigen expression (CD73, CD90, and CD105). Periosteal sheets grew faster in the novel culture medium than in the conventional medium. However, assessment of cell shape and ALP activity revealed that the periosteal cells growing in the novel medium were relatively immature. These findings suggest that the novel culture medium featuring PRFext offers advantages by shortening the culture period and excluding possible risks associated with xeno-factors without negatively altering the activity of periosteal sheets.
Highlights
Abundant growth factors and cytokines stored in platelet granules are released from activated platelets in response to tissue injury
The cell density was maximum in the central region in cultures with MSC-PCM + 2% platelet-rich fibrin extract (PRFext) (C), while the lowest density was observed in the cultures with conventional Medium199 + 10% fetal bovine serum (FBS) (A)
At the beginning of the second phase, we developed and patented a new expansion method using stocked human platelet-rich plasma (PRP) along with recombinant human basic fibroblast growth factor (bFGF) to allow the growth of periosteal sheets [12]
Summary
Abundant growth factors and cytokines stored in platelet granules are released from activated platelets in response to tissue injury. These soluble factors are involved in wound healing and tissue repair [1]. Finding a possible alternative to FBS was strongly motivated by two major reasons: (1) limitation of the variability of FBS owing to the increased demands and decreased production ability; and (2) wide variability between batches that may affect end-product reproducibility, risks of pathogen contaminations, and ethical issues [1]. The quality of PLs varied by source; shortage and risks of unexpected contamination could be avoided with the use of autologous platelets
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