Abstract
The objective of this study was to evaluate the ability of a collagen membrane (CM) as a carrier to successfully deliver platelet-derived growth factor (PDGF) and to observe the subsequent effects of the factor on preosteoblasts in vitro. MC3T3-E1 mouse preosteoblasts were cultured with a commercially available CM containing PDGF. After a two-day cell culture, cell viability was investigated by the MTT assay and cell proliferation was assessed by the crystal violet proliferation assay. Expression levels of the following osteoblastic differentiation marker genes were measured by real-time PCR: runt-related transcription factor 2 (RUNX2), osteopontin (OPN), bone sialoprotein (BSP), and osteocalcin (OCN). A cell proliferation assay was conducted, and osteoblastogenesis was determined by alkaline phosphatase (ALP) activity. A sustained release of PDGF from a CM was observed for ~3 weeks. Gene expression of all RUNX2, OPN, BSP, and OCN in CM with PDGF was significantly upregulated compared to those in CM without PDGF (all p < 0.05). Interestingly, CM without PDGF also significantly increased gene expression of RUNX2 and OPN in MC3T3-E1 cells compared to the cell control (both p < 0.05). Furthermore, it was observed that the PDGF released from CM significantly promoted ALP activity and cell proliferation with little cytotoxicity. These results suggest that a CM can be utilized for sustained delivery of PDGF. Also, released PDGF can promote MC3T3-E1 cell activities. This strategy may lead to an improvement in the current clinical treatment of bone defects in periodontal and implant therapy.
Highlights
Regenerative procedures using barrier membrane technology are presently well established in implant dentistry and periodontal therapy
Biological effects of released Platelet-derived growth factor (PDGF) from collagen membrane (CM) on cellular alkaline phosphatase (ALP) and proliferation activity To evaluate the biological effect of PDGF from CM, osteoblastogenesis was determined by ALP activity measured in MC3T3-E1 cells (Figure 2)
There was no significant difference in cell proliferation activity between CM carrying PDGF and CM alone for up to 4 days, CM carrying PDGF significantly induced cell proliferation at 6 days compared to the control CM alone (p < 0.05)
Summary
Regenerative procedures using barrier membrane technology are presently well established in implant dentistry and periodontal therapy. Collagen membranes (CMs) are frequently used in guided tissue regeneration (GTR) and guided bone regeneration (GBR), based on the premise that barrier membrane materials will promote selective cell re-population and subsequent reconstitution of the periodontal attachment apparatus as well as bone [1]. Studies have shown that recombinant human PDGF treatment of rat periodontal ligament (PDL) fibroblasts induced a strong mitogenic and chemotactic cell response, which stimulated collagen synthesis [8]. We hypothesized that a CM can be utilized as a carrier for sustained delivery of PDGF to enhance bone regeneration. This is the first study to examine the use of a commercially available CM for the delivery of PDGF. The second aim was to determine the subsequent effects of the released factor on cell viability, cell proliferation, expression levels of differentiation marker genes, and osteoblastogenesis
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