title: In vitro and in vivo biocompatibility of chitosan-xanthan polyionic complex.
abstract: A novel hydrogel, CHITOXAN(TM) (CH-X), has potential as a vehicle for controlled drug delivery. The hydrogel is obtained by complexation of two polysaccharides, chitosan and xanthan. In the present work we investigated the biocompatibility of the complex using in vitro and in vivo models. The cytotoxic effects of CH-X microspheres as well as their degradation products at different concentrations were assessed on fibroblasts (fibroblast cell line L-929) using 3-(4,5-dimethylthiazole-2yl)-2,5-triphenyl tetrazolium) (MTT). The test is based on mitochondrial dehydrogenase cell activity as an indicator of cell viability. Interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) cytokines as well as nitric oxide (NO) production by macrophages (macrophage cell line J-774) were examined as indicators of cell activation. In vivo biocompatibility assessment was performed for 1 to 12 weeks. This study was performed using tablets obtained after compression of CH-X particles implanted at the subcutaneous level in male Wistar rats. CH-X biocompatibility and degradation were investigated using histological studies. Light and transmission electron microscopy (TEM) analyses were used to determine the foreign-body reaction and phagocytosis of the implants by macrophages. Fibroblast exposition to CH-X particles and degradation products did not show cytotoxic effects as measured by MTT test. TNF-alpha production was dependent on CH-X particles concentration, whereas IL-1beta production was found to be dose independent. CH-X extract products stimulated TNF-alpha secretion when used at the highest concentration (10 mg/mL), notably after 28 days' degradation time. No effect was observed on IL-1beta production when CH-X extracts were used in comparison to the control. The effects of CH-X particles on NO secretion were similar as on TNF-alpha. Histological studies showed that CH-X tablets broke down into particles which progressively degraded into smaller fragments. A significant fraction of the fragments was ingested by the macrophages after 12 weeks of implantation. Light microscopy studies showed a weak foreign-body reaction as a function of time and the fibrous layer thickness decreased with time of implantation.
authors.affiliation: Biomedical Engineering Institute, Biomechanics and Biomaterials Research Group, Ecole Polytechnique, P.O. Box 6079, Station Down Town, Montreal, Quebec, H3C 3A7, Canada.
authors.forename: F
authors.initials: F
authors.lastname: Chellat
authors.forename: M
authors.initials: M
authors.lastname: Tabrizian
authors.forename: S
authors.initials: S
authors.lastname: Dumitriu
authors.forename: E
authors.initials: E
authors.lastname: Chornet
authors.forename: P
authors.initials: P
authors.lastname: Magny
authors.forename: C H
authors.initials: CH
authors.lastname: Rivard
authors.forename: L
authors.initials: L
authors.lastname: Yahia
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D015415:Biomarkers; D002240:Carbohydrate Sequence; D002460:Cell Line; D002686:Chitin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D048271:Chitosan; D003470:Culture Media; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D007202:Indicators and Reagents; D007375:Interleukin-1 / Q000096:biosynthesis; D008262:Macrophage Activation / Q000187:drug effects; D008297:Male; D008422:Materials Testing; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000096:biosynthesis; D010587:Phagocytosis / Q000187:drug effects; D011135:Polysaccharides, Bacterial / Q000737:chemistry* / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D001672:Biocompatible Materials; D015415:Biomarkers; D003470:Culture Media; D020100:Hydrogels; D007202:Indicators and Reagents; D007375:Interleukin-1; D011135:Polysaccharides, Bacterial; D014409:Tumor Necrosis Factor-alpha; C413466:chitoxan; D002686:Chitin; D009569:Nitric Oxide; D048271:Chitosan; C002563:xanthan gum