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   2019| May-August  | Volume 2 | Issue 2  
    Online since August 19, 2019

 
 
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ORIGINAL ARTICLE
The effects of human amniotic fluid on periodontal ligament fibroblast cell viability, proliferation, and cytokine/growth factor expression
Ahmed Gamil Ibraheem, Steven B Blanchard, Saleh Mohammed Al-Hijji, Khaled Al-Nasr-Allah, L Jack Windsor
May-August 2019, 2(2):23-29
DOI:10.4103/GFSC.GFSC_10_19  
Background: The importance of the amniotic fluid (AF) to the fetus is clear. However, very few studies have been published to examine the potential uses of this fluid in various areas such as tissue regeneration. AF contains epidermal growth factor, transforming growth factor-alpha, transforming growth factor beta-1, insulin-like growth factor-I, erythropoietin and granulocyte colony-stimulating factor, as well as hyaluronic acid and hyaluronic acid-stimulating factor. Previous studies suggest that AF can increase fibroblast proliferation and chemotaxis, and decrease apoptosis as well as promote wound healing. Furthermore, evidence showed that human AF inhibits hyaluronidase, elastase, and cathepsin. The current study examined the effects of human AF on periodontal ligament fibroblasts (PDLF) in terms of cell toxicity, cell proliferation, and cytokine/growth factor expression. Materials and Methods: Cytotoxicity of AF on PDLF was determined using lactate dehydrogenase assays. PDLF proliferation was determined using water-soluble tetrazolium-1 assays. Cytokine/growth factor expression was determined on AF-treated PDLF, AF alone, and PDLF alone utilizing protein arrays. Results: Human AF at 10% and below did not affect cell growth and was not toxic. AF-treated PDLF cells showed a decrease in cytokine/growth factor levels compared to the sum of cytokine/growth factor levels in AF only and cells only for 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased. Conclusion: Human AF at the examined concentrations was not toxic to PDLF cells and did not influence their proliferation. In addition, AF (10%) caused a decrease in the total protein levels of cytokines/growth factors expressed in 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased.
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CASE REPORT
Using concentrated growth factors as an alternative to bone graft material in sinus augmentation to rehabilitate atrophic posterior maxilla
Julio César Capella Cobos, Adolfo Enriquez Granados
May-August 2019, 2(2):30-36
DOI:10.4103/GFSC.GFSC_12_19  
A larger number of senior patients want to eliminate the use of conventional overdenture such as Conventional Complete Denture or Removable Partial Denture and replace it by an Implant-supported fixed prosthesis or at least implant-supported overdenture with the purpose of restoring and improving the masticatory function, aesthetics besides helping them gain confidence to talk and smile. Most of those patients have lost their teeth and been posteriorly edentulous for extended periods of time, causing severe bone resorption and Pneumatized Maxillary Sinus that makes it impossible for conventional dental implant placement. This report reviews two successful cases of rehabilitation of the atrophic posterior maxilla with previous bilateral ridge augmentation in Pneumatized Maxillary Sinuses using solely Concentrated Growth Factors (CGF) without bone graft augmentation with simultaneous implant placement. Where CGF is obtained from the patient's which was collected in blood collection tubes and processed by a special centrifuge device.
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TECHNICAL NOTE
Initial parameters for the assessment of osteoblast differentiation through alkaline phosphatase for biomaterial testing
Melissa Leitão, Suzana Azevedo Dos Anjos, Luciana Domenico Queiroz, Gutemberg Alves, Elena Mavropoulos
May-August 2019, 2(2):37-40
DOI:10.4103/GFSC.GFSC_6_19  
Context: MC3T3-E1 cells are preosteoblasts from mouse calvaria widely used on in vitro studies of biocompatibility under the influence of different biomaterials for bone tissue therapy, through test that includes the determination of biomarkers such as alkaline phosphatase (ALP). However, many studies use cell densities ranging from 1 × 103 to 2 × 105 cells per well even though these cells exhibit decreased proliferation when worked at high cell initial cell densities. Aims: To determine the initial parameters for the determination of ALP activity employing commercial kits and MC3T3-E1 cells for cellular differentiation studies of bone substitute biomaterials. Materials and Methods: Twenty-four well plates were seeded at concentrations of 1 × 102 to 1 × 104 at passage 8 and cultivated from 1 to 32 days. Cell density at each time was estimated through a crystal violet dye exclusion assay, cell morphology was evaluated through light microscopy, and colorimetric ALP assays were performed each day. Results: The initial cell densities of 1 × 102 and 2.5 × 102 cells per well were determined as ideal for still maintaining desired cellular characteristics for long time trials while higher initial densities demonstrated certain saturation and cell death on the 24th day. The activity of secreted ALP can be adequately measured in vitro up to 32 days after the seeding of MC3T3 cells, through commercial kits employing initial cell densities of 100–250 cells per well. Conclusion: The present protocol may provide an interesting tool for the determination of differentiation for the preclinical evauation of biomaterials.
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