RESUMO
Surface wrinkling to fabricate hierarchical surface topographies has attracted much attention because of the potential and multifunctional applications of hierarchical surface wrinkles beyond uniform wrinkles. Although many reports have described the preparation of hierarchical wrinkles induced by mechanical stress and heat, fabrication through drying-induced shrinkage has hardly been reported. Here we introduce hierarchical surface wrinkles and bumps generated on a chitosan film via the preparation of double-skin layers with κ- and ι-carrageenans, respectively, and subsequent drying. Double-skin layers are fabricated on a swollen chitosan film, called a chitosan hydrogel film, that is soaked first in κ- or ι-carrageenan solution and then in water to remove excess adsorbed κ- or ι-carrageenan. After the film is dried, hierarchical microscopic surface architectures are observed. In the case of the κ-carrageenan system, the wrinkles are hierarchical, consisting of wrinkles (6.2 ± 2.8 µm) that have smaller buckles (0.23 ± 0.09 µm). We reveal that the wrinkles or the smaller buckles are caused by plane inhomogeneous shrinkage between the κ-carrageenan layer and the chitosan film or by the aggregation of the κ-carrageenan layer upon drying, respectively. Interestingly, the ι-carrageenan system showed hierarchical bumps consisting of semispherical bumps (5.6 ± 2.1 µm) that have smaller bumps (0.78 ± 0.27 µm). We reveal that the larger bumps are generated during the immersion of the chitosan hydrogel film into ι-carrageenan solution. The smaller bumps are generated by the aggregation of the ι-carrageenan layer that occurs during drying; this process requires the plane compression strain caused by the shrinkage of the chitosan hydrogel film.
Assuntos
Quitosana , Carragenina/farmacologia , Dessecação , Estresse Mecânico , ÁguaRESUMO
The emergence of parasites resistant to praziquantel, the only therapeutic agent, and its ineffectiveness as a prophylactic agent (inactive against the migratory/juvenile Schistosoma mansoni), make the development of new antischistosomal drugs urgent. The parasite's mitochondrion is an attractive target for drug development, because this organelle is essential for survival throughout the parasite's life cycle. We investigated the effects of 116 compounds against Schistosoma mansoni cercaria motility that have been reported to affect mitochondrion-related processes in other organisms. Next, eight compounds plus two controls (mefloquine and praziquantel) were selected and assayed against the motility of schistosomula (in vitro) and adults (ex vivo). Prophylactic and therapeutic assays were performed using infected mouse models. Inhibition of oxygen consumption rate (OCR) was assayed using Seahorse XFe24 analyzer. All selected compounds showed excellent prophylactic activity, reducing the worm burden in the lungs to less than 15% of that obtained in the vehicle control. Notably, ascofuranone showed the highest activity, with a 98% reduction of the worm burden, suggesting the potential for the development of ascofuranone as a prophylactic agent. The worm burden of infected mice with S. mansoni at the adult stage was reduced by more than 50% in mice treated with mefloquine, nitazoxanide, amiodarone, ascofuranone, pyrvinium pamoate, or plumbagin. Moreover, adult mitochondrial OCR was severely inhibited by ascofuranone, atovaquone, and nitazoxanide, while pyrvinium pamoate inhibited both mitochondrial and nonmitochondrial OCRs. These results demonstrate that the mitochondria of S. mansoni are a feasible target for drug development.
Assuntos
Preparações Farmacêuticas , Esquistossomose mansoni , Esquistossomicidas , Animais , Camundongos , Mitocôndrias , Schistosoma mansoni , Esquistossomose mansoni/tratamento farmacológico , Esquistossomose mansoni/prevenção & controle , Esquistossomicidas/uso terapêuticoRESUMO
The development of anion sensors for selective detection of a specific anion is a crucial research topic. We previously reported a selective photo-induced colorimetric reaction of 1-methyl-3-(N-(1,8-naphthalimidyl)ethyl)imidazolium (MNEI) having a cationic receptor in the presence of molecules having multiple carboxy groups, such as succinate, citrate, and polyacrylate. However, the mechanism underlying this reaction was not clarified. Here, we investigate the photo-induced colorimetric reaction of N-[2-(trimethylammonium)ethyl]-1,8-naphthalimide (TENI), which has a different cationic receptor from MNEI and undergoes the photo-induced colorimetric reaction, and its analogues to clarify the reaction mechanism. The TENI analogues having substituents on the naphthalene ring provide important evidence, suggesting that the colorimetric chemical species were radical anions generated via photo-induced electron transfer from carboxylate to the naphthalimide derivative. The generation of the naphthalimide-based radical anion is verified by 1H NMR and cyclic voltammetry analyses, and photo-reduction of methylene blue is mediated by TENI. In addition, the role of the cationic receptor for the photo-induced colorimetric reaction is investigated with TENI analogues having different hydrophilic groups instead of the trimethylammonium group. Interestingly, the photo-induced colorimetric reaction is observed in a nonionic analogue having a polyethylene glycol group, indicating that the colorimetric reaction does not require a cationic receptor. On the other hand, we reveal that the trimethylammonium group stabilizes the radical anion species. These generation and stabilization phenomena of naphthalimide-based radical anion species will contribute to the development of sophisticated detection systems specific for carboxylate.
RESUMO
In order to synthesize a promising material for developing a novel peptide/protein delivery system, guanidinylation of chitooligosaccharides with 1-amidinopyrazole hydrochloride was investigated herein. The production of guanidinylated chitooligosaccharides was demonstrated by infrared spectroscopy (IR), nuclear magnetic resonance (NMR), and elemental analyses. Interestingly, we found that the reducing end in the guanidinylated chitooligosaccharides was converted to a cyclic guanidine structure (2-[(aminoiminomethyl)amino]-2-deoxy-d-glucose structure). This reaction was carefully proven by the guanidinylation of d-glucosamine. Although this is not the first report on the synthesis of the 2-[(aminoiminomethyl)amino]-2-deoxy-d-glucose, it has provided a rational synthetic route using the high reactivity of the reducing end. Furthermore, we found that the interaction between chitooligosaccharides and bovine serum albumin is weak when in a neutral pH environment; however, it is significantly improved by guanidinylation. The guanidinylated chitooligosaccharides are useful not only for the development of a novel drug delivery system but also as a chitinase/chitosanase inhibitor and an antibacterial agent.
Assuntos
Quitina/análogos & derivados , Guanidina/metabolismo , Soroalbumina Bovina/metabolismo , Animais , Bovinos , Quitina/química , Quitina/metabolismo , Quitosana , Ciclização , Guanidina/química , Estrutura Molecular , Oligossacarídeos , Oxirredução , Ligação Proteica , Soroalbumina Bovina/químicaRESUMO
Malaria is one of the three major global health threats. Drug development for malaria, especially for its most dangerous form caused by Plasmodium falciparum, remains an urgent task due to the emerging drug-resistant parasites. Exploration of novel antimalarial drug targets identified a trifunctional enzyme, malate quinone oxidoreductase (MQO), located in the mitochondrial inner membrane of P. falciparum (PfMQO). PfMQO is involved in the pathways of mitochondrial electron transport chain, tricarboxylic acid cycle, and fumarate cycle. Recent studies have shown that MQO is essential for P. falciparum survival in asexual stage and for the development of experiment cerebral malaria in the murine parasite P. berghei, providing genetic validation of MQO as a drug target. However, chemical validation of MQO, as a target, remains unexplored. In this study, we used active recombinant protein rPfMQO overexpressed in bacterial membrane fractions to screen a total of 400 compounds from the Pathogen Box, released by Medicines for Malaria Venture. The screening identified seven hit compounds targeting rPfMQO with an IC50 of under 5 µM. We tested the activity of hit compounds against the growth of 3D7 wildtype strain of P. falciparum, among which four compounds showed an IC50 from low to sub-micromolar concentrations, suggesting that PfMQO is indeed a potential antimalarial drug target.
Assuntos
Inibidores Enzimáticos/farmacologia , Malária Cerebral/tratamento farmacológico , Malária Falciparum/tratamento farmacológico , Oxirredutases/antagonistas & inibidores , Animais , Antimaláricos/metabolismo , Antimaláricos/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Malária Cerebral/enzimologia , Malária Cerebral/parasitologia , Malária Falciparum/enzimologia , Malária Falciparum/parasitologia , Malatos/metabolismo , Camundongos , Mitocôndrias/enzimologia , Oxirredutases/genética , Plasmodium berghei/efeitos dos fármacos , Plasmodium berghei/patogenicidade , Plasmodium falciparum/enzimologia , Plasmodium falciparum/patogenicidade , Quinonas/metabolismoRESUMO
The alternative oxidase (AOX) is a monotopic diiron carboxylate protein which catalyzes the four-electron reduction of dioxygen to water by ubiquinol. Although we have recently determined the crystal structure of Trypanosoma brucei AOX (TAO) in the presence and absence of ascofuranone (AF) derivatives (which are potent mixed type inhibitors) the mechanism by which ubiquinol and dioxygen binds to TAO remain inconclusive. In this article, ferulenol was identified as the first competitive inhibitor of AOX which has been used to probe the binding of ubiquinol. Surface plasmon resonance reveals that AF is a quasi-irreversible inhibitor of TAO whilst ferulenol binding is completely reversible. The structure of the TAO-ferulenol complex, determined at 2.7â¯Å, provided insights into ubiquinol binding and has also identified a potential dioxygen molecule bound in a side-on conformation to the diiron center for the first time.
Assuntos
Proteínas Mitocondriais/química , Oxirredutases/química , Oxigênio/química , Proteínas de Plantas/química , Proteínas de Protozoários/química , Trypanosoma brucei brucei/enzimologia , Ubiquinona/análogos & derivados , Cumarínicos/química , Proteínas Mitocondriais/metabolismo , Oxirredutases/metabolismo , Oxigênio/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Protozoários/metabolismo , Ressonância de Plasmônio de Superfície , Ubiquinona/química , Ubiquinona/metabolismoRESUMO
Chitins obtained by fermentation of shrimp wastes using Lactobacillus brevis with and without further inoculations with Rhizopus oligosporus resulted in higher molecular weight than the commercial biopolymer. After grinding in acidic conditions, the attained chitins were fully fibrillated by a mechanical treatment throughout ten passes in a high-pressure water jet system as evidenced by field-emission scanning electron microscopy. The chitin sample crystallinities decreased from 85% to 68%. A previous chitin sample bleaching, as well as the sonication of chitin nanofiber suspensions, enhanced the transparency in the resulting nanofiber sheets. Suspensions and sheets of chitin extracted by L. brevis with successive R. oligosporus inoculations displayed higher transmittance and acetylation degree, as well as improved mechanical properties compared to chitin extracted with only L. brevis. Mechanical studies demonstrated that Young's modulus of the nanofibers using this biological chitin was remarkably higher than that for the commercial product, an important characteristic in polymer reinforcements.
Assuntos
Exoesqueleto/química , Quitina/química , Nanofibras/química , Animais , Quitina/isolamento & purificação , Levilactobacillus brevis , Tamanho da Partícula , Propriedades de Superfície , ResíduosRESUMO
In this study, the effects of chitosan and surface-deacetylated chitin nanofibrils (SDACNFs) on hair growth were evaluated. In human follicle dermal papilla cells in vitro, chitosan and SDACNFs were shown to increase cell growth on day 3 after the initiation of treatment, together with an increase in the production of fibroblast growth factor-7 (FGF-7) by these cells on day 3. Furthermore, in an in vivo study in mice, chitosan and SDACNF application promoted hair growth. The number of anagen follicles significantly increased compared with that in the control group, whereas the number of telogen follicles significantly decreased in the chitosan and SDACNF groups. In the chitosan and SDACNFs groups, moreover, the expression levels of FGF-7 and Sonic hedgehog were significantly upregulated in hair follicles. Overall, our results demonstrated that chitosan and SDACNFs promoted hair growth and therefore may have applications as novel therapeutic agents for the treatment of hair loss in patients.
Assuntos
Quitina/farmacologia , Quitosana/farmacologia , Cabelo/crescimento & desenvolvimento , Nanofibras/química , Acetilação , Animais , Proliferação de Células/efeitos dos fármacos , Cabelo/citologia , Cabelo/efeitos dos fármacos , Folículo Piloso/citologia , Folículo Piloso/efeitos dos fármacos , Proteínas Hedgehog/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Camundongos Endogâmicos C57BL , Propriedades de SuperfícieRESUMO
We introduce a simple method for producing guanidinylation of chitosan (CS) with 1amidinopyrazole hydrochloride (AP). Production of GCS is proved via NMR, IR, and elemental analyses. In a reaction using 4.5â¯eq of AP for 7â¯days at room temperature, we obtained GCS with 42.3% of degree of guanidinylation (DG). The DG value can be controlled by the reaction time and AP amount. Furthermore, remarkable enhancement of the interaction between GCS and bovine serum albumin by guanidinylation was observed. This simple guanidinylation method for CS could provide novel additives for protein/peptide delivery systems.
Assuntos
Arginina/química , Peptídeos Penetradores de Células/química , Quitosana/química , Guanidina/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Soroalbumina Bovina/químicaRESUMO
Human dihydroorotate dehydrogenase (HsDHODH) is a key enzyme of pyrimidine de novo biosynthesis pathway. It is located on the mitochondrial inner membrane and contributes to the respiratory chain by shuttling electrons to the ubiquinone pool. We have discovered ascofuranone (1), a natural compound produced by Acremonium sclerotigenum, and its derivatives are a potent class of HsDHODH inhibitors. We conducted a structure-activity relationship study and have identified functional groups of 1 that are essential for the inhibition of HsDHODH enzymatic activity. Furthermore, the binding mode of 1 and its derivatives to HsDHODH was demonstrated by co-crystallographic analysis and we show that these inhibitors bind at the ubiquinone binding site. In addition, the cytotoxicities of 1 and its potent derivatives 7, 8, and 9 were studied using human cultured cancer cells. Interestingly, they showed selective and strong cytotoxicity to cancer cells cultured under microenvironment (hypoxia and nutrient-deprived) conditions. The selectivity ratio of 8 under this microenvironment show the most potent inhibition which was over 1000-fold higher compared to that under normal culture condition. Our studies suggest that under microenvironment conditions, cancer cells heavily depend on the pyrimidine de novo biosynthesis pathway. We also provide the first evidence that 1 and its derivatives are potential lead candidates for drug development which target the HsDHODH of cancer cells living under a tumor microenvironment.
RESUMO
The Journal of Functional Biomaterials Editorial Office have been made aware that some parts of the article [1] are duplicated from other publications[...].
RESUMO
Plasmodium falciparum is an apicomplexan parasite that causes the most severe malaria in humans. Due to a lack of effective vaccines and emerging of drug resistance parasites, development of drugs with novel mechanisms of action and few side effects are imperative. To this end, ideal drug targets are those essential to parasite viability as well as absent in their mammalian hosts. The mitochondrial electron transport chain (ETC) of P. falciparum is one source of such potential targets because enzymes, such as L-malate:quinone oxidoreductase (PfMQO), in this pathway are absent humans. PfMQO catalyzes the oxidation of L-malate to oxaloacetate and the simultaneous reduction of ubiquinone to ubiquinol. It is a membrane protein, involved in three pathways (ETC, the tricarboxylic acid cycle and the fumarate cycle) and has been shown to be essential for parasite survival, at least, in the intra-erythrocytic asexual stage. These findings indicate that PfMQO would be a valuable drug target for development of antimalarial with novel mechanism of action. Up to this point in time, difficulty in producing active recombinant mitochondrial MQO has hampered biochemical characterization and targeted drug discovery with MQO. Here we report for the first time recombinant PfMQO overexpressed in bacterial membrane and the first biochemical study. Furthermore, about 113 compounds, consisting of ubiquinone binding site inhibitors and antiparasitic agents, were screened resulting in the discovery of ferulenol as a potent PfMQO inhibitor. Finally, ferulenol was shown to inhibit parasite growth and showed strong synergism in combination with atovaquone, a well-described anti-malarial and bc1 complex inhibitor.
Assuntos
Membranas Mitocondriais/enzimologia , Oxirredutases/metabolismo , Plasmodium falciparum/enzimologia , Proteínas de Protozoários/metabolismo , Antimaláricos/farmacologia , Atovaquona/farmacologia , Biocatálise/efeitos dos fármacos , Cumarínicos/farmacologia , Sinergismo Farmacológico , Inibidores Enzimáticos/farmacologia , Humanos , Malária Falciparum/parasitologia , Malária Falciparum/prevenção & controle , Malatos/metabolismo , Membranas Mitocondriais/efeitos dos fármacos , Ácido Oxaloacético/metabolismo , Oxirredutases/antagonistas & inibidores , Plasmodium falciparum/efeitos dos fármacos , Proteínas de Protozoários/antagonistas & inibidoresRESUMO
We previously developed bio-based wrinkled surfaces induced by wood-mimetic skins upon drying in which microscopic wrinkles were fabricated on a chitosan (CS) film by immersing it in a phenolic acid solution, followed by horseradish peroxidase (HRP)-catalyzed surface reaction and drying. However, the detailed structure of the resulting wood-mimetic skins, including crosslinking mode and thickness, has not been clarified due to the difficulty of the analysis. Here, we prepare wrinkled films using ferulic acid (FE), vanillic acid (VA), and homovanillic acid (HO) and characterize their structures to clarify the unknown characteristics of wood-mimetic skin. Chemical and structural analyses of wood-mimetic skins prepared using VA and HO indicate that the crosslinking structure in the skin is composed of ionic bonds between CS and an oligophenolic residue generated by the HRP-catalyzed reaction on the CS surface. Moreover, the quantity of these ionic bonds is related to the skin hardness and wrinkle size. Finally, SEM and TOF-SIMS analyses indicate that the skin thickness is on the submicron order (<200nm).
Assuntos
Materiais Biomiméticos , Quitosana/química , Peroxidase do Rábano Silvestre/metabolismo , Catálise , MadeiraRESUMO
The development of chitin-based materials with favorable mechanical properties and biocompatibility is an important research goal owing to the wide-ranging practical applications. In this study, a composite film was prepared using chitin nanofibers and gelatin. The CNF/gelatin composite film was highly viscous and had a fine nanofiber structure. The transmittances indicated high transparency, regardless of nanofiber content. The water content of the CNF/gelatin composite film increased linearly as the gelatin content increased. Although the CNF/gelatin composite film did not induce severe inflammation, it strongly induced fibroblast proliferation, indicating high biocompatibility. Based on these results, the films are suitable for biological applications, e.g., tissue engineering, medicines, and cosmetics.
Assuntos
Materiais Biocompatíveis/química , Quitina/química , Gelatina/química , Nanofibras/química , Animais , Feminino , Gelatina/ultraestrutura , Teste de Materiais , Fenômenos Mecânicos , Camundongos , Propriedades de SuperfícieRESUMO
This study investigated the prophylactic effects of orally administered surface-deacetylated chitin nanofibers (SDACNFs) and chitosan against 5-fluorouracil (5-FU)-induced intestinal mucositis, which is a common side effect of 5-FU chemotherapy. SDACNFs and chitosan abolished histological abnormalities associated with intestinal mucositis and suppressed hypoproliferation and apoptosis of intestinal crypt cells. These results indicate that SDACNF and chitosan are useful agents for preventing mucositis induced by anti-cancer drugs.
Assuntos
Quitina/administração & dosagem , Quitina/uso terapêutico , Quitosana/uso terapêutico , Fluoruracila/efeitos adversos , Mucosite/induzido quimicamente , Mucosite/tratamento farmacológico , Nanofibras/química , Acetilação , Administração Oral , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Quitina/farmacologia , Quitosana/administração & dosagem , Quitosana/farmacologia , Feminino , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Antígeno Ki-67/metabolismo , Camundongos Endogâmicos C57BL , Nanofibras/administração & dosagem , Nanofibras/ultraestrutura , Peroxidase/metabolismoRESUMO
We previously developed biobased wrinkled surfaces based on wood mimetic skins in which microscopic wrinkles were fabricated on a chitosan film by immersion in a phenolic acid solution, horseradish peroxidase-catalyzed surface reaction, and drying. Here, we prepared a diverse range of wrinkled films by immersion treatment at 30, 40, 50, and 60 °C in p-coumaric acid and then investigated the correlation between wrinkle morphology and mechanical properties. Wrinkle wavelengths gradually decreased as the immersion temperature increased as well as the previous report. In order to clarify the mechanisms responsible for the different wrinkle morphologies, the films were subjected to elastic moduli measurement and GPC analysis after immersion treatment. These experiments provided evidence that the chitosan around the film surface decomposed along with the immersion process. The decomposition was accelerated by higher immersion temperature, suggesting that higher temperatures led to the formation of softer skins, inducing smaller wrinkles. In fact, wrinkle morphologies with this system were predominately determined by the hardness of the wood mimetic skins. This phenomenon is consistent with the fundamentals of surface wrinkling in nature. This study is the first to demonstrate that artificial wrinkling triggered by water evaporation can be controlled by precise control of the surface hardness of soft material.
Assuntos
Materiais Biomiméticos , Dessecação , Fenômenos Mecânicos , Estresse Mecânico , Propriedades de Superfície , Temperatura , MadeiraRESUMO
A protein/CaCO3/chitin nanofiber complex was prepared from crab shells by a simple mechanical treatment with a high-pressure water-jet (HPWJ) system. The preparation process did not involve chemical treatments, such as removal of protein and calcium carbonate with sodium hydroxide and hydrochloric acid, respectively. Thus, it was economically and environmentally friendly. The nanofibers obtained had uniform width and dispersed homogeneously in water. Nanofibers were characterized in morphology, transparency, and viscosity. Results indicated that the shell was mostly disintegrated into nanofibers at above five cycles of the HPWJ system. The chemical structure of the nanofiber was maintained even after extensive mechanical treatments. Subsequently, the nanofiber complex was found to improve the growth of tomatoes in a hydroponics system, suggesting the mechanical treatments efficiently released minerals into the system. The homogeneous dispersion of the nanofiber complex enabled easier application as a fertilizer compared to the crab shell flakes.
Assuntos
Exoesqueleto/química , Carbonato de Cálcio/química , Quitina/química , Nanofibras/química , Proteínas/química , Animais , Braquiúros/metabolismo , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/crescimento & desenvolvimento , Microscopia Eletrônica de Varredura , Nanofibras/toxicidade , Nanofibras/ultraestrutura , Desenvolvimento Vegetal/efeitos dos fármacos , Espectroscopia de Infravermelho com Transformada de Fourier , Estresse MecânicoRESUMO
Esterification with maleic anhydride significantly improved the mechanical disintegration of chitin into uniform 10-nm nanofibers. Nanofibers with 0.25° of esterification were homogeneously dispersed in basic water due to the carboxylate salt on the surface. Esterification proceeded on the surface and did not affect the relative crystallinity. A cast film of the esterified chitin nanofibers was highly transparent, since the film was free from light scattering.
Assuntos
Quitina/química , Anidridos Maleicos/química , Nanofibras/química , Animais , Braquiúros , Esterificação , Luz , Fenômenos Mecânicos , Nanofibras/ultraestrutura , Nanotecnologia/métodos , Espalhamento de Radiação , Propriedades de Superfície , Água/químicaRESUMO
We evaluated the effect of chitin nanofibril (CNF) application via skin swabs on an experimental atopic dermatitis (AD) model. AD scores were lower, and hypertrophy and hyperkeratosis of the epidermis were suppressed after CNF treatment. Furthermore, inflammatory cell infiltration in both the epidermis and dermis was inhibited. CNFs also attenuated histological scores. The suppressive effects of CNFs were equal to those of corticosteroid application; however, chitin did not show these effects. CNF application might have anti-infllammatory effects via suppression of the activation of nuclear factor-kappa B, cyclooxygenase-2, and inducible nitric oxide synthase. In an early-stage model of experimental AD, CNFs suppressed AD progression to the same extent as corticosteroids. They also suppressed skin inflammation and IgE serum levels. Our findings indicate that CNF application could aid in the prevention or treatment of AD skin lesions.
Assuntos
Quitina/uso terapêutico , Dermatite Atópica/terapia , Nanofibras , Animais , Quitina/química , Quitina/farmacologia , Inflamação/terapia , Camundongos , Pele/efeitos dos fármacosRESUMO
We previously reported a chitin nanofiber hydrogel from squid pen ß-chitin by a simple NaOH treatment. In the present study, a calcium phosphate/chitin nanofiber hydrogel was prepared for bone tissue engineering. Calcium phosphate was mineralized on the hydrogel by incubation in a solution of diammonium hydrogen phosphate solution followed by calcium nitrate tetrahydrate. X-ray diffractometry and Fourier transform infrared spectroscopy showed the formation of calcium phosphate crystals. The morphology of the calcium phosphate crystals changed depending on the calcification time. After mineralization, the mechanical properties of the hydrogel improved due to the reinforcement effect of calcium phosphate crystal. In an animal experiment, calcium phosphate/chitin nanofiber hydrogel accelerated mineralization in subcutaneous tissues. Morphological osteoblasts were observed.