Microencapsulated ciliary neurotrophic factor: physical properties and biological activities.
Controlled drug release in the CNS and PNS is still an obstacle to the treatment of neurodegenerative disorders. We have prepared a variety of microspheres containing either ciliary neurotrophic factor (CNTF) or genetically engineered cells able to synthesize and release this cytokine. CNTF is a multifunctional cytokine that can regulate the survival and differentiation of many types of developing and adult neurons. However, when given in therapeutically effective doses by systemic injections, it produces numerous adverse side effects. In order to minimize these effects we have microencapsulated it in biopolymers (chitosans, alginates, and copolymers in various proportions to achieve different kinetic properties). Size distribution profiles were determined by an image analysis system and surface characteristics were assessed by electron microscopy. The total content of CNTF as well as the amounts released per day were determined by ELISA and in vitro bioassays. The results from the release kinetics demonstrate that long-term secretion (up to 24 days) of CNTF is achieved by combining chitosan with copolymerized lactic and glycolic acid, whereas microspheres made of alginate provided only relatively short-term release (2-12 days). Neuron survival and neurite outgrowth in cultures of ciliary ganglia were supported by microencapsulated CNTF, indicating biological stability of CNTF. Genetically engineered human kidney cells 293 continued synthesizing CNTF within spheres and the released amounts of CNTF in the culture medium were comparable to the amounts secreted from monolayers (1 ng/ml of supernatant from confluent cultures) or even higher. These studies provide a basis for future testing of CNTF in encapsulated preparations using animal models of neurodegenerative disorders.
Maysinger
D
D
Department of Pharmacology and Therapeutics, McGill University, Montreal, Canada.
Krieglstein
K
K
Filipovic-Grcic
J
J
Sendtner
M
M
Unsicker
K
K
Richardson
P
P
D000464:Alginates; D000818:Animals; D001672:Biocompatible Materials; D001681:Biological Assay; D002214:Capsules; D002461:Cell Line, Transformed / Q000378:metabolism; D017690:Cell Transplantation / Q000379:methods; D002642:Chick Embryo; D020934:Ciliary Neurotrophic Factor; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D019344:Lactic Acid; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D009414:Nerve Growth Factors / Q000008:administration & dosage / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000008:administration & dosage* / Q000378:metabolism / Q000502:physiology*; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D011995:Recombination, Genetic
D000464:Alginates; D001672:Biocompatible Materials; D002214:Capsules; D020934:Ciliary Neurotrophic Factor; D004337:Drug Carriers; D006603:Hexuronic Acids; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D020723:Glucuronic Acid