Freeze-Dried Platelet-Rich Plasma Accelerates Bone Union with Adequate Rigidity in Posterolateral Lumbar Fusion Surgery Model in Rats

Yasuhiro Shiga, Sumihisa Orita, Go Kubota, Hiroto Kamoda, Masaomi Yamashita, Yusuke Matsuura, Kazuyo Yamauchi, Yawara Eguchi, Miyako Suzuki, Kazuhide Inage, Takeshi Sainoh, Jun Sato, Kazuki Fujimoto, Koki Abe, Hirohito Kanamoto, Masahiro Inoue, Hideyuki Kinoshita, Yasuchika Aoki, Tomoaki Toyone, Takeo Furuya, Masao Koda, Kazuhisa Takahashi, Seiji Ohtori, Yasuhiro Shiga, Sumihisa Orita, Go Kubota, Hiroto Kamoda, Masaomi Yamashita, Yusuke Matsuura, Kazuyo Yamauchi, Yawara Eguchi, Miyako Suzuki, Kazuhide Inage, Takeshi Sainoh, Jun Sato, Kazuki Fujimoto, Koki Abe, Hirohito Kanamoto, Masahiro Inoue, Hideyuki Kinoshita, Yasuchika Aoki, Tomoaki Toyone, Takeo Furuya, Masao Koda, Kazuhisa Takahashi, Seiji Ohtori

Abstract

Fresh platelet-rich plasma (PRP) accelerates bone union in rat model. However, fresh PRP has a short half-life. We suggested freeze-dried PRP (FD-PRP) prepared in advance and investigated its efficacy in vivo. Spinal posterolateral fusion was performed on 8-week-old male Sprague-Dawley rats divided into six groups based on the graft materials (n = 10 per group): sham control, artificial bone (A hydroxyapatite-collagen composite) -alone, autologous bone, artificial bone + fresh-PRP, artificial bone + FD-PRP preserved 8 weeks, and artificial bone + human recombinant bone morphogenetic protein 2 (BMP) as a positive control. At 4 and 8 weeks after the surgery, we investigated their bone union-related characteristics including amount of bone formation, histological characteristics of trabecular bone at remodeling site, and biomechanical strength on 3-point bending. Comparable radiological bone union was confirmed at 4 weeks after surgery in 80% of the FD-PRP groups, which was earlier than in other groups (p < 0.05). Histologically, the trabecular bone had thinner and more branches in the FD-PRP. Moreover, the biomechanical strength was comparable to that of autologous bone. FD-PRP accelerated bone union at a rate comparable to that of fresh PRP and BMP by remodeling the bone with thinner, more tangled, and rigid trabecular bone.

Figures

Figure 1
Figure 1
(A) FD-PRP preparation. FD-PRP appears as powder, which can be dissolved in distilled water with the same concentration of fresh PRP. (B) The artificial bone is crushed into powder. (C) FD-PRP is mixed with the powdered artificial bone followed by activation using thrombin and calcium chloride before use. (D) Schema of the spine (transplantation site). The graft material was implanted over the transverse processes of L4–L6.
Figure 2. Histological image of the lumbar…
Figure 2. Histological image of the lumbar spine (hematoxylin and eosin stain).
(A) Dashed line: horizontal line at the lower edge of the dural sac. We have measured the entire remodeling part above the dotted line, including the upper part of the vertebral arch and the spinous process. (B) Arrowheads indicate the trabecular bone branches.
Figure 3. Mechanical strength evaluation: three-point bending…
Figure 3. Mechanical strength evaluation: three-point bending test.
(A) Harvested lumbar spine (L4–L6). (B) Three-point bending. (C) Representative plotting for initial peak pressure measurement.
Figure 4. Anteroposterior radiographs of the spinal…
Figure 4. Anteroposterior radiographs of the spinal segment of the platelet-rich plasma group 4 weeks after the surgery.
(A) Sham group. (B) Artificial bone–alone group. (C) Autologous bone group. (D) Artificial bone + fresh-PRP group. (E) Artificial bone + FD-PRP group. (F) Artificial bone + BMP group. Arrowheads indicate the part of bone union, and arrows show the transverse process shadow. The FD-PRP group showed greater bone formation compared with the other groups.
Figure 5
Figure 5
(A) Evaluation of bone union. The FD-PRP and BMP groups showed earlier bone formation compared with the artificial bone–alone and autologous bone groups. (B) Amount of bone formation 8 weeks after the surgery. n.s., no significance. The FD-PRP group showed more bone formation (p < 0.05), comparable to the fresh-PRP and BMP groups.
Figure 6. Histological images of trabecular bone.
Figure 6. Histological images of trabecular bone.
(A) Artificial bone–alone group. (B) Autologous bone group. (C) Artificial bone + fresh-PRP group. (D) Artificial bone + FD-PRP group. (E) Artificial bone + BMP group. Trabecular bone formation of the FD-PRP group consisted of a tangled structure with more thin branches, compared with the autologous bone group. The trabecular bone formation is similar to that of the fresh-PRP and BMP groups.
Figure 7
Figure 7
(A) Total area ratio: the FD-PRP group was slightly inferior to the autologous bone group. (B) Number of trabecular branches: the FD-PRP group had almost the same results as the fresh-PRP and BMP groups but had more branches compared with the artificial bone–alone and autologous bone groups. (C) Quantified value of the trabecular width. The trabecular branches in the FD-PRP group formed significantly thinner branches compared with those in autologous bone groups.
Figure 8. Mechanical strength evaluation using three-point…
Figure 8. Mechanical strength evaluation using three-point bending test.
n.s. no significance. The FD-PRP group was significantly stronger than the sham group and the artificial bone–alone group (p 
All figures (8)

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Source: PubMed

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