In vitro effects of combined and sequential delivery of two bone growth factors

Human fetal osteoblast-like cells formed a regular multilayered structure in vitro with an extensive collagen-based extracellular matrix. With colloidal gold immunocytochemistry, labels for alkaline phosphatase and osteocalcin were distributed in a relatively diffuse pattern, in contrast to the bone growth factors, insulin-like growth factors I and II (IGF-I and IGF-II), transforming growth factor-beta 1 (TGF-beta 1), and basic fibroblast growth factor, which were colocalized in the collagenous matrix of the multilayer. The inclusion of 17 beta-estradiol (10(-11) to 10(-9) M) in the culture medium increased multilayer depths, increased labeling for IGF-I, IGF-II, and TGF-beta 1, and resulted in earlier detection of TGF-beta 1 label. In contrast, the increase in multilayer depth resulting from treatment with human platelets, an exogenous source of growth factors, was not accompanied by an increase in matrix IGF-I, IGF-II, or TGF-beta 1 label, suggesting a particular effect of estradiol to facilitate this process. Because growth factors in bone matrix may act as coupling agents when released during resorption, reduced growth factor incorporation in the presence of reduced sex steroid concentrations may lead to uncoupling of resorption and subsequent formation.

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Controlled Release of Bone Growth Factors from Injectable, Biodegradable Polymer Scaffolds for Bone Tissue Engineering

MRS Proceedings ◽

10.1557/proc-662-nn3.7 ◽

2000 ◽

Vol 662 ◽

Author(s):

Elizabeth L. Hedberg ◽

Antonios G. Mikos

Keyword(s):

Growth Factors ◽

Controlled Release ◽

Bone Growth ◽

Bone Ingrowth ◽

Release Kinetics ◽

Salt Content ◽

Burst Effect ◽

Plga Microparticles ◽

Model Drug

AbstractThe objective of this research is to fabricate injectable, polymeric composites that will act as scaffolds for bone ingrowth as well as carriers for the controlled release of bone growth factors. To that end, the injectable polyester poly(propylene fumarate) (PPF) was loaded with poly(DLlactic-co-glycolic acid) (PLGA) microparticles carrying the model drug FITC-dextran. This preparation was then crosslinked with N-vinyl pyrrolidinone in the presence of benzoyl peroxide as initiator and sodium chloride (NaCl) as leachable porogen. The encapsulation of growth factors in microparticles is necessary to minimize their denaturation during scaffold crosslinking. PLGA microparticles (0.04 g microparticles/g PPF) were incorporated into PPF composites having variable NaCl weight percents (50 and 70 wt% NaCl) and the effect on FITC-dextran release kinetics was determined in vitro for cylinders of diameter 6.5 mm and height 13.0 mm. The FITC-dextran loaded microparticles alone exhibited a large initial burst effect, while the composite materials displayed a smaller burst effect and a longer linear region of release. At day 3, 54.6±2.1%, 5.1±0.9%, and 12.5±0.3% of loaded FITC-dextran was released into pH 7.4 phosphate buffered saline from the microparticles, the 50 wt% NaCl, and the 70 wt% NaCl composites, respectively. By day 28, 90.9±6.9%, 12.7±1.7%, and 34.4±0.4% of loaded FITC-dextran was released. Our results demonstrate that PLGA microparticles can be incorporated into PPF composites and that the release kinetics of FITC-dextran can be systematically manipulated through alteration of the composite initial salt content.

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