Clin Orthop Surg.  2019 Sep;11(3):361-368. 10.4055/cios.2019.11.3.361.

Platelet-Rich Plasma Pretreatment on Grit-Blasted Titanium Alloy for Enhanced Osteogenic Differentiation of Human Adipose-Derived Stem Cells

• Hong SH
• Nam J
• Kim HJ
• Yoo JJ

• KMID: 2462567
• DOI: http://doi.org/10.4055/cios.2019.11.3.361

Abstract

BACKGROUND
Adequate bone formation around titanium alloy implants is integral to successful implantation surgery. Stem cell-coated implants may accelerate peri-implant bone formation. This study investigates the effect of platelet-rich plasma (PRP) pretreatment on a titanium-alloy surface in terms of proliferation and osteogenic differentiation of human adipose-derived stem cells (hADSCs).
METHODS
Allogenic leukocyte-depleted PRP was obtained from blood supernatants. The hADSCs were isolated from thigh subcutaneous fat tissue. Grit-blasted titanium plugs were used in two different groups. In one group, 200 µL of PRP was added to the grit-blasted titanium plugs. The hADSCs were seeded in two groups: grit-blasted titanium plugs with or without PRP. The number of hADSCs was measured after 4 hours, 3 days, and 7 days of culture using Cell Counting Kit-8. Osteogenesis of hADSCs was measured by using an alkaline phosphatase activity assay on days 7 and 14, and a calcium assay on days 14 and 21. Osteogenic gene expression was measured by using reverse transcription polymerase chain reaction analysis of alkaline phosphatase, osteocalcin, and type I collagen mRNA. The microscopic morphology of grit-blasted titanium plugs with or without PRP was examined with a field-emission scanning electron microscope using a JSM-7401F apparatus on days 1 and 7.
RESULTS
Proliferation and osteogenic differentiation of hADSCs were found to be significantly higher on the grit-blasted titanium alloy preprocessed with PRP than the same alloy without pretreatment. Furthermore, a structural fibrillar mesh developed compactly on the grit-blasted titanium alloy with the PRP pretreatment.
CONCLUSIONS
Our results demonstrate that a hADSC-based approach can be used for tissue-engineered peri-implant bone formation and that PRP pretreatment on the grit-blasted titanium alloy can improve proliferation and osteogenic differentiation of hADSCs.

Keyword

Platelet-rich plasma; Titanium alloy; Stem cell

MeSH Terms

Alkaline Phosphatase
Alloys*
Calcium
Cell Count
Collagen Type I
Gene Expression
Humans*
Osteocalcin
Osteogenesis
Platelet-Rich Plasma*
Polymerase Chain Reaction
Reverse Transcription
RNA, Messenger
Stem Cells*
Subcutaneous Fat
Thigh
Titanium*
Alkaline Phosphatase
Alloys
Calcium
Collagen Type I
Osteocalcin
RNA, Messenger
Titanium

Figure

• 

  Fig. 1 (A) Morphology of grit-blasted titanium alloy (TiGB) plug. Macroscopic image (B) and scanning electron microscope image (C) of the TiGB plug (×60 and ×1,000, respectively).

• 

  Fig. 2 Field-emission scanning electron microscope images (×1,000 magnification) of adhered human adipose-derived stem cells after 1 day and 7 days of culture on TiGB (A, B) and TiGB-PRP (C, D). (C) and (D) appeared different because fibrin network covered the raw surface of TiGB-PRP. Asterisks: adhered cells, arrow: fibrin network. TiGB: grit-blasted titanium alloy, PRP: platelet-rich plasma.

• 

  Fig. 3 Attachment and proliferation of human adipose-derived stem cells (hADSCs). (A) Attachment of hADSCs on TiGB and TiGB-PRP after 4 hours of culture. (B) Proliferation of hADSCs on TiGB and TiGB-PRP. TiGB: grit-blasted titanium alloy, PRP: platelet-rich plasma. *p < 0.05.

• 

  Fig. 4 Quantitative results of osteogenic differentiation of human adipose-derived stem cells (hADSCs) on plugs. (A) Alkaline phosphatase (ALP) activities of hADSCs after 7 and 14 days of culture. (B) Calcium content of hADSCs after 14 and 21 days of culture. TiGB: grit-blasted titanium alloy, PRP: platelet-rich plasma. *p < 0.05.

• 

  Fig. 5 Expression of alkaline phosphatase (ALP), osteocalcin, and type I collagen mRNA after osteogenic differentiation for 14 days (A) and 21 days (B). GAPDH: glyceraldehyde-3-phosphate dehydrogenase, TiGB: grit-blasted titanium alloy, PRP: platelet-rich plasma.

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