Bone marrow mesenchymal stem cells, platelet-rich plasma and nanohydroxyapatite-type I collagen beads were integral parts of biomimetic bone substitutes for bone regeneration

Abstract

Platelet rich plasma (PRP), which includes many growth factors, can activate osteoid production, collagen synthesis and cell proliferation. Nanohydroxyapatite-type I collagen beads (CIB), which mimetic natural bone components, are not only flexible fillers for bone defect but also encourage osteogenesis. Bone marrow mesenchymal stem cells (BMSCs) are often used as an abundant cell source for tissue engineering. We used a rabbit model to combine PRP, CIB and BMSCs (CIB+PRP+BMSC) into a bone-like substitute to study its impact on bone regeneration, when compared to defect alone, PRP, CIB+PRP, and PRP+BMSC. CIB+PRP upregulated more alkaline phosphatase (ALP) activity in BMSCs than PRP alone at 4 weeks postoperation. CIB+PRP+BMSC and PRP+BMSC did not differ significantly in DNA content, total collagen content, and ALP activity at 8 weeks. In histological assay, both CIB+PRP+BMSC and PRP+BMSC showed more bone regeneration at 4 and 8 weeks. Higher trabecular bone volume in tissue volume (BV/TV) (31.15±2.67% and 36.93±1.01%), fractal dimension (FD) (2.30±0.18 and 2.65±0.02) and lower trabecular separation (Tb.Sp) (2.30±0.18 and 1.35±0.16) of CIB+PRP+BMSC than of other groups at 4 and 8 weeks, and approach to of bone tissue (BV/TV=24.35±2.13%; FD=2.65±0.06; Tb.Sp=4.19±0.95). CIB+PRP+BMSC significantly enhanced new bone formation at 4 week. Therefore, nanohydroxyapatite-type I collagen beads combined with PRP and BMSCs produced a bone substitute with efficiently improved bone regeneration that shows promise to repair bone defects.

The Effect of Autologous Platelet-Rich Plasma on Bone Regeneration by Autologous Mesenchymal Stem Cells Loaded onto Allogeneic Cancellous Bone Granules

“…Because it is prepared by intraoperative centrifugation from whole blood on the day of surgery, and poses no risk of disease transmission, PRP has gained considerable attention over the past decade [Kasten et al, 2008]. However, there have been conflicting reports about the effect of PRP on bone regeneration [Kassolis et al, 2000;Choi et al, 2004;Fennis et al, 2004;Li et al, 2004;Roldán et al, 2004;Schlegel et al, 2004;Yamada et al, 2004;Raghoebar et al, 2005;Thorwarth et al, 2005;Plachokova et al, 2007;Kasten et al, 2008;Batista et al, 2011;Ohba et al, 2012a;Parizi et al, 2012;Shafiei-Sarvestani et al, 2012;El Backly et al, 2013;Lin et al, 2013]. In the present study, we hypothesized that additional application of autologous PRP may promote bone regeneration in transplantation of allogeneic cancellous bone granules loaded with autologous BM-MSC.…”

“…However, those studies included numerous different factors that could have influenced the results and most did not include a contralateral control group in the same experimental animal, which complicated evaluation of the effect of PRP. Various species, e.g., NWR, Japanese white rabbits [Ohba et al, 2012a], Mongrel dogs [Choi et al, 2004], and Danish Landrace pigs [Li et al, 2004] weight of the NWR have also differed [Kasten et al, 2008;Batista et al, 2011;Parizi et al, 2012;El Backly et al, 2013;Lin et al, 2013]. Baseline platelet counts vary markedly within and among species [ Table 6 in Kahn et al, 2006], and the difference in the metabolism of small and large animals may influence the rate of bone healing [Mooren et al, 2010].…”

“…In addition, various materials such as coral species [Guillemin et al, 1987;Parizi et al, 2012], deproteinized bovine bone material [Hämmerle et al, 1997], nanohydroxyapatite/poly(ester urethane) [El Backly et al, 2013], and β-tricalcium phosphate [Li et al, 2004] have been used as bone regeneration scaffolds. MSC were used in a few preclinical studies on the effect of PRP on bone regeneration [Yamada et al, 2004;Kasten et al, 2008;El Backly et al, 2013;Lin et al, 2013]. …”

“…One study in rabbits reported that 0.15 mL PRP with a mean of 807.564 ± 211.490 platelets × mm -3 from 4 mL blood with a mean of 320.133 ± 42.323 platelets × mm -3 was extracted using a 2-step technique of centrifugation at 2,400 rpm for 8 min and at 2,400 rpm for 5 min [Ohba et al, 2012a]. Another study presented that a mean PRP of 104.53 ± 47.67 × 10 7 plate- 336 lets/ml from 9.8 mL blood of a mean of 28.03 ± 13.34 × 10 7 platelets/mL was collected by 1-step centrifugation at 1,200 rpm for 10 min [Lin et al, 2013]. In addition, a PRP of 3 × 10 6 /μL was obtained using 3-step centrifugation twice at 209 g for 16 min, and at 1,500 g for 12 min [El Backly et al, 2013].…”

“…PRP has been applied to bone defects after gel preparation in many studies [Kassolis et al, 2000;Choi et al, 2004;Yamada et al, 2004;Plachokova et al, 2007;Ohba et al, 2012a;El Backly et al, 2013;Lin et al, 2013]. Because platelet gel can act as a biologic adhesive and barrier, and function as a sealant [Kassolis et al, 2000], it can maintain the PRP concentration at the defect site, further promoting bone regeneration even more.…”