BackgroundCellulose and its derivatives such as carboxymethyl cellulose (CMC) have been employed as a biomaterial for their diverse applications such as tissue engineering, drug delivery and other medical materials. Porosity of the scaffolds has advantages in their applications to tissue engineering such as more cell adhesion and migration leading to better tissue regeneration. After synthesis of CMC-poly(ethylene oxide) (PEO) hydrogel by mixing the solutions of both CMC-acrylate and PEO-hexa-thiols, fabrication and evaluation of a CMC-PEO gel and its film in porous form have been made for its possible applications to tissue regeneration. Physicochemical and biological properties of both CMC-PEO hydrogel and porous films have been evaluated by using physicochemical assays by SEM, FTIR and swelling behaviors as well as in vitro assays of MTT, Neutral red, BrdU, gel covering and tissue ingrowth into the pores of the CMC-PEO gel films. Degradation of CMC-PEO hydrogel was also evaluated by treating with esterase over time.ResultsChemical grafting of acrylate to CMC was verified by analyses of both FTIR and NMR. CMC-PEO hydrogel was obtained by mixing two precursor polymer solutions of CMC-acrylate and PEO-hexa-thiols and by transforming into a porous CMC-PEO gel film by gas forming of ammonium bicarbonate particles. The fabricated hydrogel has swollen in buffer to more than 6 times and degraded by esterase. The results of in vitro assays of live and dead, MTT, BrdU, Neutral red and gel covering on the cells showed excellent cell compatibility of CMC-PEO hydrogel and porous gel films. Furthermore the porous films showed excellent in vitro adhesion and migration of cells into their pore channels as observed by H&E and MT stains.ConclusionsBoth CMC-PEO hydrogel and porous gel films showed excellent biocompatibility and were expected to be a good candidate scaffold for tissue engineering.
BackgroundHyaluronic acid (HA) is of importance due to its diverse applications in pharmaceuticals and medical devices such as dermal filler, adhesion barriers, carrier for cells and bioactive molecules as well as scaffold biomaterials for tissue engineering. Evaluations of purification and biocompatibility of HA are required for its applications to biomaterials.ResultsAfter synthesizing HA by fermentation of streptococcus zooepidemicus for 25 hr, extensively purification of the fermented broth was performed to remove impurities using a filtration process for insoluble components and cells, and diverse adsorbents for soluble impurities. Its in vitro biocompatibility has been evaluated by measurement of cell counting and assay of cell live and dead. 60% yield of white HA powder was obtained, having 15–17 dL/g intrinsic viscosity with a molecular weight of approximately 1,000 kDa. While low molecular weight impurities and insoluble impurities were successfully removed using a ultrafiltration membrane with 50 KDa molecular weight cut, endotoxins, high molecular weight proteins and nucleic acids were removed from the broth by employing adsorbents such as alumina and activated carbons. Alumina showed the best results for the removal of endotoxins, all of the activated carbons were very effective in the removal of high molecular weight proteins and nucleic acids. The purified HA solution showed excellent cell compatibility with no cell damages as observed by both measurement of cell proliferation and observation of cell viability.ConclusionsWe obtained high molecular weight HA with excellent biocompatibility as judged by both measurement of cell proliferation and viability, indicating high possibility of its applications to biomaterials.
Design of micro-patterning of hydrogel is of critical importance in both understanding cellular behaviors and mimicking controlled microenvironments and architectures of diverse well-organized tissues. After micro-patterning of hyaluronic acid (HA) hydrogel on a poly(dimethyl siloxane) substrate, its physical and biological properties have been compared with those of a non-patterned hydrogel for its possible applications in bone tissue engineering. The micro-patterned morphologies of HA hydrogel in both swollen and dehydrated forms have been observed with light microscope and scanning electron microscope, respectively, before and after in vitro cell culture. When MC3T3 bone cells were in vitro cultured on both HA hydrogels, the micro-patterned one shows excellence in cell proliferation and lining for 7 days along the micro-pattern paths over those of the non-patterned one, which have shown less cell-adhesiveness. The cytotoxicity of the micro-patterned HA hydrogels was in vitro evaluated by the assays of MTT, BrdU and Neutral red. The viability and morphology of MC3T3 cells on both HA hydrogels were observed with a fluorescence microscope by the live & dead assay, where their viability was confirmed by staining of F-actin development. The results of their H&E staining showed that both micro-patterned and non-patterned hydrogels induced development of tissue regeneration as observed by cell attachment, proliferation, and survivability, but the micro-patterned one induced distinctive patterning of both better initial cells adhesion on the micro-patterns and subsequently development of their proliferation and extracellular matrix, which were considered as important characteristics in their applications to tissue engineering.
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