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. 2021 Nov 25;26(23):7131.
doi: 10.3390/molecules26237131.

Lyophilised Platelet-Rich Fibrin: Physical and Biological Characterisation

Nurul Aida Ngah  1   2 , George J Dias  3 , Darryl C Tong  1 , Siti Noor Fazliah Mohd Noor  4 , Jithendra Ratnayake  1 , Paul R Cooper  1 , Haizal Mohd Hussaini  1   5
Affiliations

Lyophilised Platelet-Rich Fibrin: Physical and Biological Characterisation

Nurul Aida Ngah et al. Molecules. .

Abstract

Background: Platelet-rich fibrin (PRF) has gained popularity in craniofacial surgery, as it provides an excellent reservoir of autologous growth factors (GFs) that are essential for bone regeneration. However, the low elastic modulus, short-term clinical application, poor storage potential and limitations in emergency therapy use restrict its more widespread clinical application. This study fabricates lyophilised PRF (Ly-PRF), evaluates its physical and biological properties, and explores its application for craniofacial tissue engineering purposes.

Material and methods: A lyophilisation method was applied, and the outcome was evaluated and compared with traditionally prepared PRF. We investigated how lyophilisation affected PRF's physical characteristics and biological properties by determining: (1) the physical and morphological architecture of Ly-PRF using SEM, and (2) the kinetic release of PDGF-AB using ELISA.

Results: Ly-PRF exhibited a dense and homogeneous interconnected 3D fibrin network. Moreover, clusters of morphologically consistent cells of platelets and leukocytes were apparent within Ly-PRF, along with evidence of PDGF-AB release in accordance with previously reports.

Conclusions: The protocol established in this study for Ly-PRF preparation demonstrated versatility, and provides a biomaterial with growth factor release for potential use as a craniofacial bioscaffold.

Keywords: craniofacial regeneration; lyophilisation; platelet concentrate; platelet-rich fibrin; tissue engineering.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Platelet-rich fibrin (PRF) preparation. (A) Venous blood centrifuged for separation. PRF clot formed in the tube centre between red blood cells and platelet-poor plasma fractions. Appearance of (B) fresh PRF, (C) frozen PRF, and (D) lyophilised PRF.
Figure 2
Figure 2
Physical appearance of lyophilised PRF. Ly-PRF (A) before and (B) after being ground into granules using a mortar and pestle.
Figure 3
Figure 3
Representative scanning electron microscope (SEM) micrographs of surface of intact Ly-PRF, which showed irregular-surface topographical appearance. SEM magnifications: ((A) ×45, (B) ×180, (C) ×200, (D) ×270, (E) ×350, and (F) ×650). (Bar = 100 µm).
Figure 4
Figure 4
Cross-sections of intact Ly-PRF demonstrated a highly porous structure with a mixture of homogenous and heterogenous pore sizes. (SEM; original magnification: (A) ×130, (B) ×230, (C) ×600, (D) ×170, (E) ×100 and (F) ×200). (Bar = 100 µm) Images (A,B) from Donor 1, images (C,D) from Donor 2 and images (E,F) from Donor 3.
Figure 5
Figure 5
Highly dense and interconnected 3D fibrin microarchitecture in cross-sections of intact Ly-PRF. (SEM; original magnification: (A) ×1700, (B) ×1400, (C) ×1300, (D) ×2000, (E) ×1,000 and (F) ×1300). (Bar = 10 µm). (A,B) Donor 1; (C,D) Donor 2; (E,F) Donor 3.
Figure 6
Figure 6
Representative SEM micrographs of cross-sections of intact Ly-PRF revealed entrapped clusters of platelets (yellow arrows) and leukocytes (red circles) within the Ly-PRF clot. (A) Representative image of a zone of enriched platelets with various degrees of activation. (SEM; original magnification: (A,B) ×1800, (C) ×1000, (D) ×2000, (E) ×1000 and (F) ×1800). (Bar = 10 µm).
Figure 7
Figure 7
Cumulative kinetics for PDGF-AB growth factor release from five donors samples. Data presented as mean ± SD of five biological replicates (n = 5) with 2 technical replicates (n = 2).
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