Papers by Author: John P. LeGeros

Abstract: The osteoconductive property of calcium phosphate (CaP) biomaterials allow attachment, proliferation, migration, and phenotypic expression of bone cells leading to formation of new bone. The purpose of research is to develop new method of mineralizing commercial GBR membranes with calcium phosphate (CaP) and determine cell response. Resolut Adapt LT (Gore-tex) composed of co-polymer PGA/TMC and Biomend Extend (Zimmer) composed of bovine collagen were used. Membranes were mineralized with CaP using precipitation and new microwave methods. The mineralized and non-mineralized membranes were characterized using SEM, EDS, XRD, FT-IR, and TGA. Cell response to mineralized and non-mineralized membranes was determined using human osteoblastlike cells (MG-63). Microwave method was more efficient in terms of amount of minerals incorporated with membranes and time required. SEM, EDS, and FT-IR identified carbonate apatite in the mineralized membranes. No significant difference in cell proliferation was observed between mineralized and non-mineralized membranes. Greater production of type 1 collagen was observed with CaP mineralized membranes.

Abstract: The purpose of this study was to determine the effect of sintering conditions on microporosity of and cell proliferation and bone ingrowth on biphasic calcium phosphate (BCP) bioceramics. Discs were prepared from a calcium-deficient apatite preparation that upon sintering at 1050oC and above, results in a BCP with 60% hydroxyapatite (HA)/ 40% beta-tricalcium phosphate (β-TCP) ratio. The discs were divided into groups which were sintered under different conditions of heating rate (programmed vs. non-programmed) and temperature (1050°C vs. 1200°C). The discs were characterized in terms of composition (HA/β-TCP ratio), surface morphology, surface area, surface microporosity, per cent microporosity, and dissolution properties. The in vitro effect of sintering conditions on cell proliferation was determined using an established mouse fibroblast cell line (L929). Results demonstrated the following: (a) the HA/β-TCP ratio remained 60/40 regardless of sintering conditions; (b) the % microporosity, surface microporosity, surface area of the BCP and cell proliferation on the BCP significantly decreased with increasing sintering temperature, and (c) the extent of dissolution decreased with decreasing per cent microporosity. The in vivo study indicated no tissue adverse reaction and direct bone contact with the implant surface, confirming the biocompatibility of the BCP bioceramics. Resorption of the BCP and bone ingrowth was directly related to the sintering temperature: the higher the temperature, the lower the resorption and the bone ingrowth. Results of this study indicate that per cent microporosity of the BCP bioceramics affects its dissolution properties and cell response. The study demonstrates that optimum per cent microporosity elicits optimum cell response and should be considered to provide osteogenic/osteoinductive property to bioceramics.

Abstract: Since 1952 when Branemark first reported osseointegration of titanium (Ti) with bone, many academic and industrial research activities have endeavored to improve the efficacy of Ti or Ti alloy (Ti6Al4V) by modifying the chemistry, topography and design of the implant surface. Strong bonding between implant and host tissue minimize the micromovements that promote fibrous tissue formation at the implant interface that may lead to implant failure. Surface design include lateral holes perpendicular to the implant axis, grooves, variations of spacings between ridges, etc. Physico-mechanical means of surface modification is by grit-blasting with various abrasives (alumina, silica, apatitic abrasive), laser ablation, spark discharge, etc. Chemical modifications include: acid etching, treatment with alkali, treatment with fluoride, coating with titanium or with calcium phosphate (by plasma spray, electromagnetic sputtering, electrochemical deposition). A review of studies on Ti or Ti alloy implants with different surfaces showed the following methods to enhance osseointegration and greater bone formation: (1) grit-blasting with apatitic abrasive; (2) acid-etching with mixed acids; (3) adjusting plasma-spray parameter to get a higher HA/ACP ratio in the coating; (4) employing electrochemical deposition (with pulse modulation) or precipitation to obtain thin coating with homogeneous composition; and/or (5) Ftreatment.

Abstract: Osteoporosis is a ‘silent’ disease characterized by thinning cortical bone and disorganized trabecular architecture causing bone fragility leading to fracture. Osteoporosis results when the rate of bone resorption far exceeds the rate of bone formation. Current pharmaceutical interventions (estrogen therapy, bisphosphonate-based drugs) focus on inhibiting bone resorption. However, some of these therapies have serious side effects (e.g., cancer risk from estrogen therapy; osteonecrosis of the jaw and delayed fracture healing from bisphosphonate-based drugs). The long term objective of the study was to develop a novel material for potential osteoporosis therapy, prevention and fracture repair. This novel material MZF-CaP or synthetic bone mineral, SBM) incorporates Mg, Zn and F ions in a calcium phosphate matrix. Separately, magnesium (Mg), zinc (Zn) and fluoride (F) ions have been associated with biomineralization and osteoporosis therapy in human and in animals. MZF-CaP or SBM was prepared by a modified hydrolysis method previously described and characterized using x-ray diffraction, FT-IR spectroscopy, inductive coupled plasma and dissolution in acidic buffer. Separately, male and female Sprague-Dawley rats were randomly assigned to the following groups depending on the diet: GA: normal on basic diets; GB: on mineral deficient diets (md); GC: on md + Mg-CaP; GD: on md + Zn-CaP; GE: md+F-CaP; and GF: md+MZF-CaP. The rats were sacrificed after 3 months and the femur bones separated, cleaned of extraneous soft tissues and stored until needed for analyses. Femur bones were analyzed using microradiography (Faxitron), scanning electron microscopy (SEM) and microCT. Results: SEM, Faxitron and microCT analyses showed thinning of cortical bone and disorganized trabecular bone architecture for osteoporotic rats on mineral deficient diet (GB) and prevention of bone loss in rats receiving the supplemented diets (GC,GD,GE,GF). Conclusion: These results indicate that the novel material, MZF-CaP or SBM had a potential for osteoporosis therapy and prevention. Studies to demonstrate the use of SBM in reversing (recovering) bone loss are in progress.

Abstract: Plasma-sprayed ‘HA’ coatings on commercial orthopedic and dental implants were developed to combine the strength of the metal (Ti or Ti alloy) and the bioactivity of the hydroxyapatite (HA). Several studies have shown that ‘HA’-coated implants provided greater amount of bone attachment, higher bone-implant interfacial strength and accelerated skeletal attachment. However, some reports on implant failures have been attributed to coating delamination and coating early resorption of the plasma sprayed ‘HA’ coating. This paper reviews studies on characterization and degradation of plasma-sprayed ‘HA’ coatings on orthopedic and dental implants and offers alternatives to plasma-spray method of providing calcium phosphate coating. X-ray diffraction analyses showed that plasma-sprayed HA coating consists principally of HA and amorphous calcium phosphate (ACP) with minor amounts of other resorbable calcium phosphates (α- or β-tricalcium phosphates, tetracalcium phosphate), sometimes calcium oxide. The HA/ACP ratios were found to range from 20HA/80ACP to 70HA/30ACP in coated implants from different manufacturers. In vitro initial dissolution rates in acidic buffer (pH 6, 37oC) increased with decreasing HA/ACP ratios in the coating because of the preferential dissolution of the ACP phase. These results suggest that coating with very low HA/ACP ratio may result in the premature resorption of the coating before the bone can attach to the implant thus causing loosening and eventual failure of the implant. Alternatives to plasma-sprayed ‘HA’ are implant surface modifications and low temperature calcium phosphate coatings using electrochemical deposition method or precipitation method.

Abstract: Fluoride, when incorporated in the apatite, stabilizes the structure. The purpose of this study was to determine the consequences of fluoride (F) substitution on the physico-chemical properties of apatites. F-containing apatites were prepared by precipitation or by hydrolysis of CaHPO4 in solutions containing different F concentrations and characterized using x-ray diffraction, FT-IR spectroscopy, scanning electron microscopy, thermogravimetry and chemical analyses. Results showed that F incorporation have the following effects: (a) decrease in a-axis dimension, (b) increase in crystal size and thickness, (c) decrease in calcium deficiency, and (d) lower solubility.

Abstract: Plasma-sprayed HA coating combines the strength of the metal and the bioactivity of the HA. However, this method has several disadvantages. Alternatives to the plasma-spray method such as electrochemical deposition (ECD) and biomimetic or precipitation methods are being explored. The purpose of this study was to develop an ECD method for coating Ti alloy substrate with different calcium phosphates (octacalcium phosphate, calcium deficient apatite, carbonatesubstituted apatite, fluoride-substituted apatite). Pairs of Ti6Al4V plates that have been mechanically polished, ultrasonically cleaned, acid etched, rinsed and dried were used as anodes and cathodes. ECD was carried out using programmed pulse time electric fields. Results showed that uniform coating with only the desired calcium phosphate can be obtained using metastable calcium phosphate solutions at different pH and temperature conditions and different electrolyte concentrations. Coating thickness varied with the duration of coating deposition. Crystal size varied with other ECD conditions (e.g., pulse time, current density). This method can be used to obtain uniform coating of the desired calcium phosphate composition at low temperatures (25 to 80oC) on substrates of any type of geometry.

Abstract: Our earlier studies showed that several ions inhibit the crystal growth of apatite and promote the formation of amorphous calcium phosphates (ACP). These ions include: magnesium (Mg), zinc (Zn), stannous (Sn), ferrous (Fe), carbonate (CO3), pyrophosphate (P2O7). The purpose of this study was to investigate the effect of combination of these ions (e.g., Mg & CO3, Mg & P2O7, Mg & Zn, etc) on the formation and stability of ACP. ACP compounds containing the different ions were prepared at 25 and 37oC according to the method we previously described. Chemical stability was investigated by suspending the different ACP preparations in solutions with or without inhibitory ions. Thermal stability was determined by sintering the ACP at different temperatures. Dissolution properties were determined in acidic buffer. The ACP before and after chemical or thermal treatment were analyzed using X-ray diffraction, infrared spectroscopy, and thermogravimetry. Results showed synergistic effects of inhibitory ions on the formation of ACP. ACP materials, regardless of their composition, remained amorphous even after heat treatment at 400oC. Transformation of ACP to other calcium phosphate phases depended on the pH and on the solution composition.