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ORIGINAL RESEARCH REPORT
Bone forming ability of recombinant human collagen peptide
granules applied with β-tricalcium phosphate fine particles
Tomokazu Furihata
1
| Hirofumi Miyaji
1
| Erika Nishida
1
| Akihito Kato
1
|
Saori Miyata
1
| Kanako Shitomi
1
| Kayoko Mayumi
1
| Yukimi Kanemoto
1
|
Tsutomu Sugaya
1
| Tsukasa Akasaka
2
1
Department of Periodontology and
Endodontology, Faculty of Dental Medicine,
Hokkaido University, Sapporo, Japan
2
Department of Biomedical Materials and
Engineering, Faculty of Dental Medicine,
Hokkaido University, Sapporo, Japan
Correspondence
Hirofumi Miyaji, Department of
Periodontology and Endodontology, Faculty of
Dental Medicine, Hokkaido University, N13,
W 7, Kita-ku, Sapporo 060-8586, Japan.
Email: miyaji@den.hokudai.ac.jp
Funding information
AMED Translational Research Program, Grant/
Award Numbers: A105, A154; JPSP KAKENHI,
Grant/Award Numbers: JP16K11822,
JP18K17038
Abstract
Recombinant human collagen peptide, developed based on human collagen type I,
contains an arginyl-glycyl-aspartic acid (RGD)-rich motif to enhance cell behavior and
is anticipated as a xeno-free polymer material for use in tissue engineering. We fabri-
cated granules containing recombinant human collagen peptide (RCP) applied with
beta-tricalcium phosphate fine particles (RCP/β-TCP) as bone filling scaffold material
and assessed the bone forming ability of RCP/β-TCP. Recombinant peptide was ther-
mal crosslinked and freeze-dried to prepare RCP. An aqueous dispersion of β-TCP
fine particles was added to RCP to obtain RCP/β-TCP. Subsequently, RCP/β-TCP
were characterized using scanning electron microscopy (SEM), energy dispersive X-
ray spectrometry (EDX), and cell culture assessments. Furthermore, RCP/β-TCP were
implanted into rat cranial bone defects for radiographic and histological evaluations.
In SEM and EDX analyses of RCP/β-TCP, β-TCP particles dose-dependently covered
the surface of RCP. Cell culture tests showed that RCP/β-TCP remarkably promoted
proliferation and mRNA expression of various genes, such as integrin β1 and osteo-
genic markers, of osteoblastic MC3T3-E1 cells. Histomorphometric assessment at
4 weeks showed that RCP/β-TCP significantly promoted new skull bone formation
compared to RCP (p< 0.05) and control (no application) (p< 0.01). Accordingly, these
findings suggest RCP/β-TCP possess bone forming capability and would be beneficial
for bone tissue engineering therapy.
KEYWORDS
bone filling material, integrin β1, osteogenic differentiation, rat skull, recombinant peptide
based on human collagen type I
1|INTRODUCTION
Improvement of bone tissue engineering therapy is required for the
treatment of bone loss caused by infectious disease, trauma, and can-
cer. Three elements of tissue engineering are proposed (O'Keefe &
Mao, 2011); cells (Diederichs et al., 2010; Fawzy El-Sayed et al., 2015;
Perez et al., 2018), signaling molecules (Kim, Lee, & Kim, 2018;
Peticone et al., 2017; Yin et al., 2018), and natural and artificial
scaffolds (Carrel et al., 2016; Nathanael et al., 2017; Xing et al., 2013)
are essential for bone conductive strategies. Scaffolds play a major
role in stimulating cell proliferation and differentiation and providing
growth and nutrition factors in bone defects (Bose, Roy, &
Bandyopadhyay, 2012). The polymer matrix is widely known as a
good bioabsorbable scaffold material (Hamlet, Vaquette, Shah,
Hutmacher, & Ivanovski, 2017; Sheikh et al., 2016). Especially, colla-
gen type I has great cellular affinity and biodegradability as scaffold
Received: 16 February 2020 Revised: 15 April 2020 Accepted: 18 April 2020
DOI: 10.1002/jbm.b.34632
J Biomed Mater Res. 2020;108B:3033–3044. wileyonlinelibrary.com/journal/jbmb © 2020 Wiley Periodicals, Inc. 3033