Bone forming ability of recombinant human collagen peptide granules applied with β‐tricalcium phosphate fine particles

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 fabricated 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 thermal 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 osteogenic 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.

Periodontal Tissue Regeneration by Recombinant Human Collagen Peptide Granules Applied With β-Tricalcium Phosphate Fine Particles

“…In this project, β-TCP was added to promote bone formation through ion supply, but was adjusted to a submicron size to ensure smooth resorption. As a result, no residual β-TCP was observed histologically, which is consistent with previous reports [30] and suggests that it does not produce long-term effects on tissue replacement and periodontal regeneration.…”

“…Accordingly, the replacement of β-TCP/RCP appears to be slower than that of the beta-TCP-blended bovine collagen scaffold previously reported by Ogawa et al [25]. The slow resorption of β-TCP/RCP was likely caused by the non-connected pore structure (calf-derived collagen sponge has a fully connected pore structure) and a difference in the degree of cross-linking [30]. However, there was almost no accumulation of CD3-positive cells to β-TCP/RCP, suggesting that there is no concern about detrimental inflammatory reactions against periodontal tissue regeneration caused by residual β-TCP/RCP [51].…”

“…These results suggest that β-TCP/RCP implantation promoted bone healing. Furihata et al reported that co-culture of β-TCP/RCP and osteoblast-like cells promoted mRNA expression of the osteogenic markers runt-related transcription factor 2 (Runx2), alkaline phosphatase, and bone sialoprotein, to simultaneously stimulate integrin β1 expression [30]. Integrin-mediated cell adhesion to extracellular matrix components, such as type I collagen, is known to activate focal adhesion kinase and its downstream target, bone morphogenetic protein (BMP)-smad signaling, which is involved in promoting osteoblast differentiation [31].…”

“…Based on these studies, Furihata et al prepared RCP granules with β-TCP fine particles (β-TCP/RCP) and showed a significant promotion of osteoblast growth and differentiation, as well as in vivo (in rat) bone formation, compared to normally granulated RCP (without β-TCP) [30]. Therefore, we hypothesized that β-TCP/RCP represents a promising scaffold material for periodontal tissue engineering.…”

A regulatory perspective on recombinant collagen-based medical devices

“…Currently, rCols have been investigated for clinical translations in many medical applications, including dermal filler [ 22 ], wound dressing [ 23 ], drug or cell carrier [ 24 , 25 ], bone void filler [ 26 ], and tissue engineering scaffolds [ 27 , 28 ]. After querying the NMPA device database [ 29 ], several rColb-MDs have been approved for market entry in China, and the statistic number of products with various expected uses is shown in Fig.…”