The Effect of Laminin-1-Doped Nanoroughened Implant Surfaces: Gene Expression and Morphological Evaluation

Abstract

Aim. This study aimed to observe the morphological and molecular effect of laminin-1 doping to nanostructured implant surfaces in a rabbit model. Materials and Methods. Nanostructured implants were coated with laminin-1 (test; dilution, 100 μg/mL) and inserted into the rabbit tibiae. Noncoated implants were used as controls. After 2 weeks of healing, the implants were removed and subjected to morphological analysis using scanning electron microscopy (SEM) and gene expression analysis using the real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Results. SEM revealed bony tissue attachment for both control and test implants. Real-time RT-PCR analysis showed that the expression of osteoblast markers RUNX-2, osteocalcin, alkaline phosphatase, and collagen I was higher (1.62-fold, 1.53-fold, 1.97-fold, and 1.04-fold, resp.) for the implants modified by laminin-1 relative to the control. All osteoclast markers investigated in the study presented higher expression on the test implants than controls as follows: tartrate-resistant acid phosphatase (1.67-fold), calcitonin receptor (1.35-fold), and ATPase (1.25-fold). The test implants demonstrated higher expression of inflammatory markers interleukin-10 (1.53-fold) and tumour necrosis factor-α (1.61-fold) relative to controls. Conclusion. The protein-doped surface showed higher gene expression of typical genes involved in the osseointegration cascade than the control surface.

Titanium Surface Coating with a Laminin-Derived Functional Peptide Promotes Bone Cell Adhesion

“…While it is known to be involved in adhesion across a broad range of cell types [2], Ln2-P3 has been chiefly evaluated in terms of its effectiveness in promoting epithelial cell attachment rather than in estimating its affinity for bone cells [10–12]. A few studies have been found to test the bone response of a laminin-coated titanium (Ti) surface [13–16]. However, the coated material has been laminin, the protein itself, not a peptide derived from it.…”

Laminin-1 acts as an adhesive for odontoblast-like cells and promotes their differentiation toward a hard tissue-forming phenotype

“…This effect was inhibited in the presence of PD98059, a specific ERK inhibitor (27), indicating that the osteogenic effect of LN-5 was at least partially mediated through the ERK signaling pathway. Moreover, a study using a rabbit model demonstrated that ALP, OCN and Runx2 were upregulated by LN-1coated nanoroughened implants (16). The differences in results might have been due to the different cell types and experimental designs employed.…”

“…The early appearance of LN-1 in tooth germ indicates that it likely has roles in the process of tooth germ development. Although it has been reported that LN is effective in promoting the adhesion and differentiation of a number of other cell types (16), its roles in the MDPC-23 cell line, a type of odontoblast-like cell, remain elusive. Importantly, the specific effects of matrix proteins depend largely on cell type and experiment design, and therefore their appropriate concentrations must be established for each specific cell type.…”

Gene expression, immunohistochemical and microarchitectural evaluation of bone formation around two implant surfaces placed in bone defects filled or not with bone substitute material

“…Finally, RT-PCR was conducted using the StepOne system (Applied Biosystems, Foster City, CA, USA) and the SybrGreen (Primer Design Ltd, Southampton, UK) fluorescent dye system under the following cyclic conditions: 95° for 10 min for primary denaturation, 35 cycles of 95° for 15 s for tape opening, and finally 60° for 60 s for annealing of the primers and extension of the fragments. The relative gene expression was calculated by reference to the expression of the mitochondrial ribosomal proteins and normalized by the gene expression of the bone fragments removed from the implants in the given period of 60 days [ 21 , 22 ].…”