The role of hypoxia in the regulation of osteogenesis and angiogenesis coupling in intraoral regenerative procedures: a review of the literature.

Intraoral bone grafting is routinely employed for implant site development. Bone graft consolidation is a complex biologic process depending on the formation of blood vessels into the augmented area. Hypoxia-inducible factors (HIFs) and hypoxia-mimicking agents (HMAs) are key stimulators of blood vessel formation. Hypoxia prevents HIFs from degradation, thus signaling angiogenesis. Under normoxia, HMAs prevent degradation of HIFs. The cellular and molecular mechanisms responsible for angiogenic-osteogenic coupling and the therapeutic manipulation of HIFs and HMAs in intraoral bone repair and regeneration are discussed. Such discoveries suggest promising approaches for the development of novel therapies to improve intraoral bone repair and regeneration procedures.

Attachment and proliferation of human osteoblast-like cells on guided bone regeneration (GBR) membranes in the absence or presence of nicotine: an in vitro study.

PURPOSE: To compare in vitro the attachment and proliferation of human osteoblast-like cells (MG63) on tissue culture plates and guided bone regeneration (GBR) membranes in the absence or presence of nicotine. MATERIALS AND METHODS: Membrane samples were fixed to wells and the cell number (CN) was counted after 24 hours (attachment assay) or 5 days (proliferation assay). The ratio of cell count (RCC) (CN at 5 days/CN at 24 hours) was calculated. The study had three parts: First, five different resorbable GBR membranes were compared (Resolut Adapt LT [RALT], Biocollagen [BC], Bio-Gide [BG], OsseoGuard [OG], and Demokritos Human Tissue Bank [DEM]). Next, cells were cultured on tissue culture plates with five different concentrations of nicotine. Finally, cells were cultured on the membrane that had demonstrated the highest RCC and CN in part 1 with four different concentrations of nicotine. RESULTS: At 24 hours, BG showed the highest CN and OG showed the lowest CN. At 5 days, BG showed the highest CN. The order of RCC was BG > OG > DEM > RALT ~ BC. At 24 hours, lower nicotine concentrations (0.3 and 3 µg/mL) showed higher CNs versus the control, whereas CNs for high nicotine concentrations (30 and 300 µg/mL) were lower than for the control. CN at 5 days and RCC were lowest with 300 µg/mL nicotine. At 24 hours and 5 days, all differences among wells with membrane were statistically insignificant. Nevertheless, CN at 5 days and RCC were highest with the lowest nicotine concentration (3 µg/mL) and lowest with high concentrations. CONCLUSIONS: Membrane materials influence attachment and proliferation of bone cells and, therefore, could affect the outcomes of GBR. On both tissue culture plates and membranes, there is a tendency toward a biphasic effect of nicotine, with stimulatory effects at low concentrations and inhibitory effects at high concentrations.

The therapeutic potential of oxygen tension manipulation via hypoxia inducible factors and mimicking agents in guided bone regeneration. A review.

Intraoral bone grafting is routinely employed for implant site development prior or simultaneously to implant placement. Bone graft consolidation is a complex biological process depending on the formation of blood vessels into the augmented area. It is highly regulated by the angiogenesis and osteogenesis coupling phenomenon. The vascular system apart from supplying nutrients and oxygen to the developing and regenerating bone, also delivers critical signals which stimulate mesenchymal cell differentiation towards an osteogenic phenotype. Hypoxia inducible factors (HIFs) and mimicking agents (HMAs) (or alternatively HIF stabilizing agents) are considered to act as key stimulators of blood vessel formation. Under normoxia, HIFs are rapidly degraded. However, their degradation is prevented under hypoxia, which in turn, triggers angiogenesis. Hence, the major role of HMAs is to prevent degradation of HIFs under normoxic conditions. Recent studies suggest that HIFs and HMAs trigger the initiation and promotion of angiogenic-osteogenic cascade events. In vitro and animal studies involving genetic manipulation of individual components of the HIFs and HMAs have provided clues to how angiogenic-osteogenic coupling is achieved. Evidence from preclinical studies further suggests that topical application of HMAs enhance angiogenesis in intraoral augmented sites. In this article, we review the current understanding of the cellular and molecular mechanisms responsible for angiogenic-osteogenic coupling. We also discuss the therapeutic manipulation of HIFs and HMAs in intraoral bone repair and regeneration. Such discoveries suggest promising approaches for the development of novel therapies to improve intraoral bone repair and regeneration procedures.

Chemical modification of an implant surface increases osteogenesis and simultaneously reduces osteoclastogenesis: an in vitro study.

OBJECTIVES: The present study investigated the effect of a chemical modification of the SLA surface (SLActive surface) on human periodontal ligament (hPDL) cell (1) adhesion, (2) proliferation, (3) osteogenic differentiation (core binding factor α-1 [Cbfa-1], bone morphogenetic protein-7 [BMP-7] gene expression and alkaline phosphatase [ALP] activity) and (4) osteoclast formation and activity (osteoprotegerin [OPG] and receptor activator of nuclear factor-κ B ligand [RANKL] gene expression). The above activities were based on the hypothesis that the expression of such molecules might be dependent on the characteristics of the implant surface. MATERIAL AND METHODS: hPDL cells were isolated and characterized for their mesenchymal origin, fibroblastic and osteoblastic phenotype. hPDL cells were cultured on smooth, SLA and SLActive implant surfaces (chemically modified). Cell attachment and proliferation were assessed for 5, 24, 72 h, 5 and 7 days. Cbfa-1, BMP-7, OPG and RANKL gene expression was assessed by RT-PCR and a colorimetric assay for ALP activity was applied. RESULTS: hPDL cells grown on SLActive surfaces demonstrated increased proliferation rates (24 h, 5 and 7 days of the incubation period), and ALP activity was found to be significantly upregulated (5, 72 h and 7 days) as compared with the SLA surfaces. After 7 days of culture, the gene expression of BMP-7, Cbfa-1 and OPG by hPDL cells was significantly upregulated, while RANKL gene expression was significantly downregulated in response to the SLActive surface. CONCLUSION: Chemical modification of a previously roughened implant surface increases hPDL proliferation and upregulates osteoblastic differentiation. It can also suppress osteoclastogenesis regulating the RANKL-RANK-OPG axis. Hence, an osteoprotective microenvironment is created around chemically modified implants that may enhance osseointegration.

Analysis of osteoblastic gene expression in the early human mesenchymal cell response to a chemically modified implant surface: an in vitro study.

OBJECTIVES: The effect of a chemical modification of the SLA surface (SLActive surface) on human bone marrow-derived mesenchymal cells (hMSCs) on; (1) adhesion, (2) proliferation and (3) early transcriptional control of osteogenic differentiation was investigated. We are based on the hypothesis that expression patterns of genes responsible for osteogenesis might be dependent on the characteristics of the implant surface. MATERIAL AND METHODS: hMSCs were allowed to grow on smooth (SMO-control), SLA and SLActive implant surfaces (chemically modified). Cell attachment and proliferation were assessed at 3 and 24 h using a MTT dye reduction assay. At 24 h of culture, DNA microarray analysis examined alterations in early gene expression using a human osteogenesis gene array, including 109 cDNAs in quadruplicates of major regulatory genes for osteogenesis. RESULTS: Initial attachment and proliferation were found to be significantly reduced. Nineteen genes were significantly upregulated when hMSCs were cultured on the SLA surfaces and 27 genes were significantly upregulated when hMSCs were cultured on the SLActive surfaces. Upregulated genes control cell differentiation, signal transduction, cell cycle regulation, angiogenesis, cell adhesion and extracellular matrix and bone formation. DISCUSSION: Chemical modification decreases further cell attachment and proliferation and upregulates early osteoblastic differentiation genes. Hence, a microenvironment is created around chemically modified implants that may enhance osseointegration.

Platelet-rich plasma effect on periodontally affected human gingival fibroblasts: an in vitro study.

OBJECTIVE: The purpose of this study was to evaluate the gingival fibroblast proliferative response derived from patients with chronic and aggressive periodontitis to homologous platelet-rich plasma (PRP). MATERIALS AND METHODS: Gingival fibroblasts derived from nine patients with chronic periodontitis, aggressive periodontitis and healthy periodontium were grown. Medium was replaced with DMEM containing 0.5% FBS in which cells remained for two days. Cells were incubated and cultured with medium containing 50 microl/ml homologous platelet-rich plasma (PRP) or not containing PRP (control) for 24 and 48 hours. PRP originated from three donors. Cell proliferation effect was evaluated at 24 and 48 hours. Cell viability was assessed with a hemocytometer. Viable cells were counted under a phase contrast microscope. RESULTS: The results revealed that incubation of human gingival fibroblasts, derived from healthy and intact periodontium, chronic periodontitis and aggressive periodontitis, in culture medium containing homologous PRP statistically significantly increased the cell proliferation at 24 and 48 hours of culture. CONCLUSION: The addition of PRP to human gingival fibroblast cultures significantly increased the proliferative response, irrespective of the presence of periodontitis, type of periodontitis and PRP donor.