James C. McPherson

Synthesis of osteotropic hydroxybisphosphonate derivatives of fluoroquinolone antibacterials

1-Hydroxybisphosphonate derivatives of ciprofloxacin, gatifloxacin and moxifloxacin have been synthesized using Cu(I) catalyzed azide-alkyne 1,3-dipolar cycloaddition reaction. The 1,2,3-triazol linked hydroxybisphosphonate derivative of ciprofloxacin exhibited antibacterial activity comparable to the parent antibiotic and all fluoroquinolone-bisphosphonates displayed osteotropic properties in a bone model.

Effect of rhBMP‐2 dose on bone formation/maturation in a rat critical‐size calvarial defect model

BACKGROUND Application of recombinant human bone morphogenetic protein-2 (rhBMP-2) has been associated with significant adverse events in craniofacial settings, including swelling and seroma formation. Recent work has demonstrated an inverse relationship between bone formation/maturation and rhBMP-2 dose, frequency/severity of adverse events increasing with rising dose. OBJECTIVE The objective of this study was to determine the most effective dose for rhBMP-2 soak-loaded onto an absorbable collagen sponge (ACS) carrier for bone formation/maturation using an established defect model. METHODS One hundred sixty-eight outbred male Sprague-Dawley rats, age 11-13 weeks, weight 325-375 g randomized into seven groups of 24 subdivided into groups of eight, were used to provide radiographic and light microscopy observations of bone formation/maturation and aberrant healing events at 2, 4 and 8 weeks following application of rhBMP-2/ACS into critical-size, ø8-mm, through-through, calvarial osteotomy defects for a dose of 1.25, 2.5, 5.0, 10.0 and 20.0 μg rhBMP-2/defect, or serve as ACS or sham-surgery controls. RESULTS rhBMP-2 dosages ≥ 2.5 μg/defect showed histological defect closure >90% within 2 weeks, and complete resolution within 4 weeks. Adverse healing events including swelling, excessive bone formation or seroma formation could not be determined with certainty in this defect model. Notably ACS control sites showed complete defect closure at the 8-week healing interval. CONCLUSIONS rhBMP-2/ACS accelerates local bone formation in the rat critical-size through-through calvarial defect model once reaching an osteoinductive dose threshold. This threshold may already be reached at a 1.25-/2.5-μg dose in this model. No further enhancement to bone formation/maturation may be observed adding rhBMP-2 above the 2.5-μg dose. The 1.25-20.0 μg dose range did not invoke appreciable aberrant healing events.

The Effect of Residual Calcium in Decalcified Freeze-dried Bone Allograft in a Critical-sized Defect in the Rattus norvegicus Calvarium

Demineralized freeze-dried bone allograft (DFDBA), a widely used graft material in periodontal regenerative procedures, is processed with hydrochloric acid in the attempt to expose proteins located within the bone matrixes that are capable of inducing new bone formation. However, the degree of DFDBA demineralization varies between tissue banks, which may have an effect on clinical regeneration. This study uses the critical-sized defect (CSD) model to evaluate the wound-healing response to the residual calcium of donor bone. If the percentage of residual calcium in a graft were demonstrated to significantly enhance wound healing, then periodontal patients may benefit from further standardization of human-allograft processing. Sixty adult, male, Harlan Sprague-Dawley rats (Rattus norvegicus) were randomly and equally divided into 4 test groups (ie, DFDBA at 1%, 2%, and 3% to 6% residual calcium levels and FDBA at 23% residual calcium) and a control group (no allograft). An 8-mm-diameter craniotomy was made in the rat calvarium, and polytetrafluoroethylene membranes with pore sizes of 0.50 microm were placed intracranially and ectocranially. Treatment materials were carefully placed into the CSD with a new sterilized dental amalgam carrier. Tetracycline hydrochloride was injected intraperitoneally for labeling new bone growth, and animals were euthanized 12 weeks postsurgery. As a result, histomorphometric bone fill at 12 weeks showed a statistically significant increase in the 2% DFDBA group as compared to all other groups. The authors conclude that a 2% residual calcium level in human DFDBA appears to significantly (P < or = .05) enhance osseous wound healing in the rat calvarium.

Effect of the Wicking Behavior of Multifilament Sutures

The purpose of this study was to compare the wicking propensity of multifilament sutures. Dexon II, Vicryl, and black silk suture (BSS) were dipped in saline or soaked for 48 h, then suspended on a microscope slide. Fluorescein isothiocyanate-dextran (FITC-D) was placed at the suture mid points, and its movement was observed using fluorescence microscopy. The experiment was repeated, replacing the FITC-D with mixture of S. salivarius and saline, incubating the suture specimens in culture medium, and evaluating microbial growth. Dipped sutures showed FITC-D movement in the Dexon II group only. All 48-h soaked sutures demonstrated FITC-D movement with significant (p < 0.005) differences in mean times: BSS 179 +/- 42 s; Vicryl 120 +/- 26 s; and Dexon II 32 +/- 2 s. Dexon II suture demonstrated wicking of S. salivarius, whereas Vicryl and BSS did not (p < 0.05). These results suggest that BSS and Vicryl sutures do not wick as readily as Dexon II does.

Histologic Evaluation of Osseous Regeneration Following Combination Therapy With Platelet-Rich Plasma and Bio-Oss in a Rat Calvarial Critical-Size Defect Model

Platelet-rich plasma (PRP) is an autogenous source of growth factors shown to facilitate human bone growth. Bio-Oss, an osteoconductive xenograft, is used clinically to regenerate periodontal defects, restore dental alveolar ridges, and facilitate sinus-lift procedures. The purpose of this study was to analyze whether a combination of PRP and Bio-Oss would enhance bone regeneration better than either material alone. PRP and/or Bio-Oss were administered in an 8-mm critical-size defect (CSD) rat calvarial model of bone defect between 2 polytetrafluoroethylene membranes to prevent soft tissue incursion. Eight weeks after the induction of the CSD, histologic sections were stained with hematoxylin and eosin stain and analyzed via light microscopy. Qualitative analyses revealed new bone regeneration in all 4 groups. The Bio-Oss and PRP plus Bio-Oss groups demonstrated greater areas of closure in the defects than the control or PRP-only groups because of the space-maintaining ability of Bio-Oss. The groups grafted with Bio-Oss showed close contact with new bone growth throughout the defects, suggesting a stronger graft. The use of PRP alone or in combination with Bio-Oss, however, did not appear to enhance osseous regeneration at 8 weeks. Areas grafted with Bio-Oss demonstrated greater space-maintaining capacity than controls, and PRP was an effective vehicle for placement of the Bio-Oss. However, at 8 weeks this study was unable to demonstrate a significant advantage of using PRP plus Bio-Oss over using Bio-Oss alone.

An In Vitro Evaluation of the Growth of Human Periodontal Ligament Fibroblasts after Exposure to a 4-META–containing Methacrylate-based Endodontic Sealer

INTRODUCTION In this study we evaluated the cytotoxic effects of MetaSEAL, a 4-META-containing meth-acrylate-based endodontic sealer, on human periodontal ligament (HPDL) fibroblasts. There are a limited number of studies on the cytotoxic effects of MetaSEAL, and there are no studies on the cytotoxic effects of MetaSEAL on cells it might come into contact with in vivo. METHODS MetaSEAL concentrations of 25, 50, 100, 200, 400, and 800 μg/mL were exposed to HPDL fibroblast cultures and evaluated at 1, 3, 7, 14, and 21 days. Controls included untreated cells and cells treated with ethanol, the vehicle for MetaSEAL suspension. Crystal violet staining in 24-well plates and the fluorescence-based CyQUANT Cell Proliferation Assay in 96-well plates assessed fibroblast viability. RESULTS Significant cytotoxicity against HPDL growth by MetaSEAL was both time- and concentration-dependent. At day 1 there were no significant cytotoxic effects, whereas by day 3, 800 μg/mL concentration, by day 7, 200, 400, and 800 μg/mL concentrations, and by day 14, 50, 100, 200, 400, and 800 μg/mL concentrations were significantly cytotoxic. By day 21, all concentrations were significantly cytotoxic. These findings were confirmed by both the crystal violet and CyQUANT assays. CONCLUSIONS MetaSEAL endodontic sealer has increasing HPDL cytotoxicity with both concentration and time exposure.