Regulating the structure of bacterial cellulose by altering the expression of bcsD using CRISPR/dCas9 / 生物工程学报

Gluconacetobacter xylinus is a primary strain producing bacterial cellulose (BC). In G. xylinus, BcsD is a subunit of cellulose synthase and is participated in the assembly process of BC. A series of G. xylinus with different expression levels of the bcsD gene were obtained by using the CRISPR/dCas9 technique. Analysis of the structural characteristics of BC showed that the crystallinity and porosity of BC changed with the expression of bcsD. The porosity varied from 59.95%-84.05%, and the crystallinity varied from 74.26%-93.75%, while the yield of BC did not decrease significantly upon changing the expression levels of bcsD. The results showed that the porosity of bacterial cellulose significantly increased, while the crystallinity was positively correlated with the expression of bcsD, when the expression level of bcsD was below 55.34%. By altering the expression level of the bcsD gene, obtaining BC with different structures but stable yield through a one-step fermentation of G. xylinus was achieved.

Traumatic perforations of the tympanic membrane: immediate clinical recovery with the use of bacterial cellulose film / Perfurações traumáticas da membrana timpânica: recuperação clínica imediata com o uso de fragmento de celulose bacteriana

Abstract Introduction: Perforation of the tympanic membrane is a reasonably frequent diagnosis in otorhinolaryngologists' offices. The expectant management is to wait for spontaneous healing, which usually occurs in almost all cases in a few weeks. However, while waiting for healing to be completed, the patients may experience uncomfortable symptoms. Although some research suggests the use of various materials to aid in the recovery of the tympanic membrane, none presented robust evidence of improvement in the cicatricial process. Nevertheless, the occlusion of the perforation with some material of specific texture and resistance can alleviate the patients' symptoms and accelerate the healing process. Objective: To evaluate the clinical (symptomatic and functional) improvement after the placement of bacterial cellulose film (Bionext®) on tympanic membrane perforations (traumatic). Methods: We evaluated 24 patients, victims of traumatic perforations of the tympanic membrane, who were evaluated in the Otorhinolaryngology Emergency Room. Following otoscopy and audiometric examination was performed, before and after the use of cellulose film occluding the tympanic membrane perforation. Results: Twenty-four patients were included, whose degree of overall discomfort caused by the tympanic membrane perforation and the presence of symptoms of autophonia, ear fullness and tinnitus were investigated. The mean score attributed to the overall annoyance caused by tympanic membrane perforation was 7.79, decreasing to a mean value of 2.25 after the film application. Symptom evaluation also showed improvement after using the film: autophonia decreased from a mean value of 6.25 to 2.08, tinnitus from 7 to 1.92 and ear fullness from 7.29 to 1.96. The auditory analysis showed mean threshold values still within the normal range at low and medium frequencies, with slight hearing loss at acute frequencies, but with significant improvement at all frequencies, with the exception of 8000 Hz, after film use. Conclusion: The use of bacterial cellulose film fragment on traumatic perforations of the tympanic membrane promoted immediate functional and symptomatic recovery in the assessed patients.

Resumo Introdução: A perfuração da membrana timpânica é uma condição clínica relativamente frequente em consultórios de otorrinolaringologistas. A conduta é quase sempre expectante, aguardando cicatrização espontânea, que costuma ocorrer na quase totalidade dos casos em algumas semanas. No entanto, enquanto não se completa, os pacientes mantêm sintomas desconfortáveis. Embora algumas pesquisas sugiram o uso de materiais diversos para auxiliar na recuperação da membrana timpânica, nenhuma apresentou evidência substancial de melhoria no processo cicatricial. Por outro lado, a oclusão da perfuração com alguns materiais de textura e resistência específicas poderia aliviar os sintomas dos pacientes durante o processo cicatricial. Objetivo: Avaliar a melhoria clínica (sintomática e funcional) após a colocação de película de celulose bacteriana (Bionext®) sobre a perfuração da membrana timpânica (traumática). Método: Foram avaliados pacientes com perfurações traumáticas da membrana timpânica que deram entrada no pronto-socorro de otorrinolaringologia. Eles foram avaliados em relação a suas queixas e exame audiométrico, antes e após a aplicação de película de celulose que oclui a perfuração da membrana timpânica. Resultados: Foram incluídos 24 pacientes, nos quais foram pesquisados o grau de incômodo global causado pela perfuração da membrana timpânica e a presença de sintomas de autofonia, plenitude auricular e tinnitus. A média da nota atribuída ao incômodo global causado pela perfuração foi de 7,79; caiu para valor médio de 2,25 após a aplicação da película. A avaliação dos sintomas também apresentou melhoria após uso da película, autofonia caiu de valor médio de 6,25 para 2,08; zumbido de 7 para 1,92 e plenitude auricular de 7,29 para 1,96. A análise auditiva apresentou um valor médio de limiares ainda dentro da normalidade em frequências baixas e médias, com perda de audição leve em frequências agudas, porém com melhoria significante em todas as frequências, com exceção de 8000 Hz, após a colocação da película. Conclusão: A aplicação de fragmento de película de celulose bacteriana sobre perfurações traumáticas da membrana timpânica promoveu recuperação funcional e sintomática imediata nos pacientes estudados.

Characterization and optimization of bacterial cellulose produced by Acetobacter spp.

im: The study aimed to search for a potential bacterial strain capable of producing maximal bacterial cellulose under optimized conditions for future scale up.Methodology: Eight cellulose producing bacterial strains were isolated from sugarcane juice using Hestrin and Schramm (HS) medium. These isolates were identified through 16S rDNA based on molecular phylogenetic approach and the cellulose mat was analyzed for their physico-chemical properties. Morphological, chemical and physical properties of cellulose mat was studied by Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy spectrum, High Performance Liquid Chromotograpy and Differential Scanning Calorimetry. Optimal nutrient composition for maximum cellulose production by the isolate was carried out by response surface methodology.Results: 16S rDNA sequence analysis revealed that the isolates belonged to Acetobacter senegalensis, Acetobacter thailandicus, Acetobacter lambici, Acetobacter lovaniensis and Acinetobacter baumannii. Among them, A. senegalensis MA1 produced the maximum cellulose mat of dry weight 3.6 g per 500 ml of HS medium after 2 weeks of incubation. The morphology of bacterial cellulose produced by A. senegalensis MA1 had a pellicle shaped distinguished network structure and the rod shaped bacterium was attached to the bottom of cellulose mat. HPLC analysis revealed that the peaks obtained from bacterial cellulose were almost similar to carboxymethyl cellulose (CMC) peak. In cellulose mat, the weight of carbon, oxygen were 60.13 and 25.12, respectively. Analysis of differential scanning calorimetry showed endothermal peak at 139.8°C., indicating the melting point of bacterial cellulose mat. Through response surface methodology analysis, 5 g of glycerol and 1.2 g of yeast extract per 100 ml medium was optimized to get maximum cellulose production (2.04 g) from A. senegalensis MA1. Interpretation: Optimized condition for maximum bacterial cellulose production may be applied at industrial scale for commercial utilization

Morphological changes in the tissue structures afeter thermal burns on the background of using dermarm wound dressing

Over the last decade, the interest in using bacterial cellulose in medicine has increased. The article publishes the data about the efficiency of healing burn wounds in rabbits in experimental conditions with the use of the DermaRM wound dressing, compared to the traditionally used Panthenol ointment and the Branolind N salve dressing

An efficient method of Yarrowia lipolytica immobilization using oil- and emulsion-modified bacterial cellulose carriers

Background: Yarrowia lipolytica is a nonconventional, dimorphic yeast with multiple biotechnological applications. Considering the size of Y. lipolytica cells and a plethora of its morphological forms (spherical cells or hyphae and pseudohyphae), it is highly difficult to select a suitable carrier for this useful microorganism. Bacterial cellulose (BC) is currently considered one of the most promising immobilization carriers. In the current study, the usefulness of oil- and emulsion-modified BCs as a carrier for Y. lipolytica immobilization was investigated. Static and agitated cultures were conducted in media supplemented with oil or emulsion to improve carrier porosity. Results: It was found that the application of oil- and emulsion-modified BCs correlated with significantly higher efficiency of Y. lipolytica immobilization and hence higher yield than the yield achieved with an unmodified carrier. Increased efficiency of immobilization correlated with BC porosity-related parameters, which, in turn, depended on the size of oil droplets introduced into the culture medium. Moreover, changes in porosity-related parameters caused by the addition of oil or emulsion to the medium were observed when the cultures were conducted only under static conditions and not under agitated conditions. Conclusion: The application of oil- and emulsion-modified BCs as carriers significantly increased the efficiency of Y. lipolytica immobilization as compared to unmodified BC. The addition of oil or emulsion to the culture medium can be a simple but effective method to modify the porosity of BC-based carriers.

Association between Vitamin D Status and Type 1 Diabetes Mellitus in Saudi Adolescents

Background: Nowadays, the deficiency of vitamin D (VD) is a health problem worldwide, that affects many people including adolescentswho have type 1 diabetes mellitus (T1DM). However, the role of VD in autoimmune diseases such as T1DM has been a recent interest. Aim:This study was designed to assess the VD deficiency prevalence in Saudi adolescents with or without T1DM. Methods: In this case-controlstudy, 49 T1DM and 49 control (non-DM) (N=98), age and gender-matched were enrolled. The study was carried out from May toSeptember 2017 at King Abdulaziz University Hospital (KAUH). After obtaining the consent form, the blood samples were withdrawn todetermine fasting blood glucose (FBG) and glycosylated hemoglobin (HbA1c) in blood and VD (25OHD) in serum. Statistical analysis wasmade by SPSS version 22. Results: Data showed that 25OHD levels were significantly lower in adolescents with T1DM compared to thecontrols (49.5 ± 26.9 nmol/L vs 67.96 ± 30.03 nmol/L). In the T1DM adolescents, VD was deficient in 44.9%, insufficient in 36.7%, andsufficient in 18.4% as compared with 34.7% (deficient), 26.5% (insufficient), and 42.86% (sufficient) in non-DM adolescents. Overall, VDwas deficient and insufficient in 81.6% of the T1DM adolescents and 61.2% of non-DM adolescents, respectively. Considering sex, femalesshowed higher significance between T1DM and non-DM groups in overall groups, and in all VD level subgroups, males showedsignificance (p<0.05) in only overall groups and in insufficient VD levels. The data showed an inverse correlation between HbA1c and FBGvalues with VD concentration. While there was no correlation between both glycemic parameters with VD in non-DM group. Conclusion:The VD deficiency prevalence in T1DM Saudi adolescents was relatively high particularly in females. Therefore, screening for VD statusand supplementation in early young age should be warranted.

A medium supplemented with vegetable extract used for cellulose production of Rhodococcus SP. MI 2 / Meio suplementado com extrato vegetal utilizado na produção de celulose de Rhodococcus SP. MI 2

In this study, the most suitable vegetable extract was screened to use as non-conventional nutrient sources for cellulose production of Rhodococcus sp. MI 2. SH medium or a synthetic medium was used as a conventional or control medium. Cha-poo (Piper sarmentosum Roxb.) and sweet potato (Ipomoea batatas Lam.) were 2 out of 14 vegetable extracts chosen as medium supplements. Rhodococcus sp. MI 2 gave the highest cellulose yield in a medium supplemented with Cha-poo extract. The optimum culture conditions in the medium supplemented with Cha-poo extract at room temperature (25o C) under static condition were 5% (v v-1) inoculum size, a 6 -day -incubation period, pH 3, 3% sucrose, and 0.5% (NH4)2SO4. The cellulose yield in the medium supplemented with Cha-poo extract was increased about 3 times (6.83 g L-1 during 6 days) higher than that obtained before optimizing (2.39 g L-1 during 6 days). The medium supplemented with Cha-poo extract cost one quarter (0.5 USD L-1 of medium) of the SH medium (1.9 USD L-1 of medium). The structure of the microfibrils of cellulose produced by Rhodococcus sp. MI 2 in a medium supplemented with Cha-poo extract observed by SEM had larger, less crowded fibrils than those produced in the medium supplemented with sweet potato extract. In addition, the microfibrils of the former had many beehive shaped knots whereas those of the latter had mantle-like surrounding the fibrils.

Neste estudo, o extrato vegetal mais adequado foi triado para uso como fontes não convencionais de nutrientes para produção de celulose de Rhodococcus sp. MI 2. Utilizou-se um meio SH ou meio sintético como meio convencional ou de controle. Cha-poo (Piper sarmentosum Roxb.) e batata-doce (Ipomoea batatas Lam.) foram 2 dos 14 extratos vegetais escolhidos como suplementos do meio. Rhodococcus sp. MI 2 deu o maior rendimento de celulose em um meio suplementado com extrato de Cha-poo. As condições ótimas de cultivo no meio suplementado com extrato de Cha-poo em temperatura ambiente (25 ºC) em condição estática foram 5% (v v-1) do tamanho do inóculo, um período de 6 dias de incubação, pH 3, 3% de sacarose, e 0,5% (NH4) 2SO4. O rendimento de celulose no meio suplementado com extrato de Cha-poo foi aumentado cerca de 3 vezes (6,83 g L-1 durante 6 dias), maior do que o obtido antes da otimização (2,39 g L-1 durante 6 dias). O meio suplementado com extrato de Cha-poo custou um quarto (0,5 USD L-1 de meio) do meio SH (1,9 USD L-1 de meio). A estrutura das microfibrilas de celulose produzidas por Rhodococcus sp. MI 2 em meio suplementado com extrato de Cha-poo, observado por MEV, apresentou fibrilas maiores e menos congestionadas do que aquelas produzidas no meio suplementado com extrato de batata-doce. Além disso, as microfibrilas do primeiro possuíam muitos nós em forma de colmeia, enquanto os do último tinham um aspecto tipo manto ao redor das fibras.

Cost-effective production of bacterial cellulose using acidic food industry by-products

Abstract To reduce the cost of obtaining bacterial cellulose, acidic by-products of the alcohol and dairy industries were used without any pretreatment or addition of other nitrogen sources. Studies have shown that the greatest accumulation of bacterial cellulose (6.19 g/L) occurs on wheat thin stillage for 3 days of cultivation under dynamic conditions, which is almost 3 times higher than on standard Hestrin and Schramm medium (2.14 g/L). The use of whey as a nutrient medium makes it possible to obtain 5.45 g/L bacterial cellulose under similar conditions of cultivation. It is established that the pH of the medium during the growth of Gluconacetobacter sucrofermentans B-11267 depends on the feedstock used and its initial value. By culturing the bacterium on thin stillage and whey, there is a decrease in the acidity of the waste. It is shown that the infrared spectra of bacterial cellulose obtained in a variety of environments have a similar character, but we found differences in the micromorphology and crystallinity of the resulting biopolymer.

Advances in the research of antibacterial composite dressings based on bacterial cellulose / 中华烧伤杂志

Bacterial cellulose (BC) is a promising material for wound dressing due to its predominant water binding capacity, mechanical property, biodegradability, and histocompatibility. Whereas BC itself exhibits no antimicrobial activity. To gain antimicrobial activity, several kinds of antibacterial agents have been introduced into BC. However, all of the antibacterial composite dressings are still in the stage of experimental research. In this paper, the types, antibacterial mechanism, and shortcomings of antibacterial composite dressings based on BC are summarized, in order to make prospects of the research trends of antibacterial composite dressings based on BC in future.

Production of Cellulose and Profile Metabolites by Fermentation of Glycerol by Gluconacetobacter Xylinus

ABSTRACT Because of the widespread occurrence of cellulose in nature, many organisms use glycerol as a source of carbon and energy, so these organisms have drawn attention to the potential use of glycerol bioconversion. The bacteria Gluconacetobacter xylinus, a strictly aerobic strain that performing incomplete oxidation of various sugars and alcohols to cellulose biosynthesis. For this reason, we modify the Hestrim-Schram medium, associating glycerol, glucose and sucrose varying their concentration. The fermentations were performed statically at a temperature of 28˚C for a period of 10 days. The pH, membrane formation, crystallinity and the production of some metabolites of the 4th, 7th and 10th days was evaluated. The results showed a higher yield of membrane in the medium containing glucose, gly 1 + glu2 on 10 fermentation of 3.5 g %. Through solid-state NMR gave the crystallinity of the membranes, where there was a clear trend toward higher crystallinity membranes with 7 days of fermentation. Metabolic products found in the media by analysis of NMR spectroscopy in liquid were similar, especially for the production of alanine and lactate that were present in all media. The leucine and threonine were present in various media, although in small quantities has been found glutamate.

Cost-effective production of bacterial cellulose using acidic food industry by-products

Abstract To reduce the cost of obtaining bacterial cellulose, acidic by-products of the alcohol and dairy industries were used without any pretreatment or addition of other nitrogen sources. Studies have shown that the greatest accumulation of bacterial cellulose (6.19 g/L) occurs on wheat thin stillage for 3 days of cultivation under dynamic conditions, which is almost 3 times higher than on standard Hestrin and Schramm medium (2.14 g/L). The use of whey as a nutrient medium makes it possible to obtain 5.45 g/L bacterial cellulose under similar conditions of cultivation. It is established that the pH of the medium during the growth of Gluconacetobacter sucrofermentans B-11267 depends on the feedstock used and its initial value. By culturing the bacterium on thin stillage and whey, there is a decrease in the acidity of the waste. It is shown that the infrared spectra of bacterial cellulose obtained in a variety of environments have a similar character, but we found differences in the micromorphology and crystallinity of the resulting biopolymer.

Bacterial cellulose and bacterial cellulose/polycaprolactone composite as tissue substitutes in rabbits' cornea / Celulose bacteriana e composto de celulose bacteriana/policaprolactone para substituto tissular na córnea de coelho

In order to test the performance of bacterial cellulose/polycaprolactone composite (BC/PCL) and pure bacterial cellulose (BC) as tissue substitutes in rabbits' cornea, a superficial ulcer containing 5mm in diameter and 0.2mm deep was made in the right cornea of 36 rabbits, then a interlayer pocket was created from the basis of this ulcer. Twelve rabbits received BC/PCL membrane and 12 were treated with BC membranes, both membranes with 8mm in diameter. The remaining rabbits received no membrane constituting the control group. The animals were clinically followed up for 45 days. Three animals of each group were euthanized at three, seven, 21, and 45 days after implantation for histological examination of the cornea along with the implant. Clinical observation revealed signs of moderate inflammatory process, decreasing from day 20th in the implanted groups. Histology showed absence of epithelium on the membranes, fibroplasia close to the implants, lymph inflammatory infiltrate with giant cells, collagen disorganization, with a predominance of immature collagen fibers in both groups with implants. Although inflammatory response is acceptable, the membranes used does not satisfactorily played the role of tissue substitute for the cornea during the study period.(AU)

Com objetivo de testar o desempenho do compósito celulose bacteriana/policaprolactona (CB/PCL) e da celulose bacteriana pura (CB) como substitutos teciduais em córnea de coelhos, foi realizada uma úlcera superficial de 5 mm de diâmetro e 0,2 mm de profundidade na córnea direita de 36 coelhos, criando-se um bolso interlamelar a partir da base dessa úlcera. Doze animais receberam a membrana do compósito CB/PCL e 12 foram tratados com membranas de CB, ambas com 8 mm de diâmetro, os coelhos restantes não receberam nenhuma membrana, constituindo o grupo controle. Os animais foram acompanhados clinicamente até 45 dias. Três animais de cada grupo sofreram eutanásia aos três, sete, 21 e 45 dias após o implante das membranas para análise histológica da córnea juntamente com o implante. À observação clínica, houve sinais de processo inflamatório moderado, diminuindo a partir do 20º dia nos grupos implantados. A histologia demonstrou ausência de epitélio sobre as membranas, fibroplasia próxima aos implantes, infiltrado inflamatório linfo-histiocitário com células gigantes, desorganização do colágeno, com predominância de fibras imaturas de colágeno em ambos os grupos com implantes. Embora a resposta inflamatória seja aceitável, as membranas utilizadas não desempenharam satisfatoriamente o papel de substituto tecidual para a córnea, no período estudado.(AU)

Treatment of tympanic membrane perforation using bacterial cellulose: a randomized controlled trial / Tratamento do tímpano perfurado com enxerto de celulose bacteriana: ensaio clínico controlado e randomizado

ABSTRACT INTRODUCTION: Promising treatments for tympanic membrane perforation closure have been studied. Therapies derived from tissue engineering probably eliminate the need for conventional surgery. Bacterial cellulose is presented as an alternative that is safe, biocompatible, and has low toxicity. OBJECTIVES: To investigate the effect on healing of direct application of a bacterial cellulose graft on the tympanic membrane compared to the conventional approach with autologous fascia. METHODS: Randomized controlled trial. Forty patients with tympanic membrane perforations secondary to chronic otitis media were included, and were randomly assigned to an experimental group (20), treated with a bacterial cellulose graft (BC) and control group (20), treated with autologous temporal fascia (fascia). We evaluated the surgical time, hospital stay, time of epithelialization and the rate of tympanic perforation closure. Hospital costs were compared. The statistical significance level accepted was established at p < 0.05. RESULTS: The closure of perforations was similar in both groups. The average operation time in the fascia group was 76.50 min versus 14.06 min bacterial cellulose in the group (p = 0.0001). The hospital cost by the Brazilian public health system was R$ 600.00 for the bacterial cellulose group, and R$ 7778.00 for the fascia group (p = 0.0001). CONCLUSION: Bacterial cellulose grafts promoted the closure of the tympanic membrane perforations, and were demonstrated to be innovative, effective, safe, minimally invasive, efficacious and to have a very low cost.

RESUMO INTRODUÇÃO: Tratamentos promissores para o fechamento da perfuração da membrana timpânica vêm sendo estudados. Terapias provenientes de engenharia de tecidos provavelmente eliminarão a necessidade de uma intervenção cirúrgica convencional. A celulose bacteriana apresenta-se como uma alternativa por ser segura, de baixa toxicidade, biocompatível. OBJETIVOS: Investigar o efeito da aplicação direta do enxerto da celulose bacteriana na cicatrização de perfurações da membrana timpânica, comparado ao procedimento convencional com fáscia autóloga. MÉTODO: Incluíram-se 40 pacientes com perfuração da membrana timpânica por otite média crônica simples. Randomizados de 1 a 40, onde os ímpares (20) foram tratados com enxerto de celulose bacteriana (CB), e os pares (20), com enxerto de fáscia temporal autóloga (fáscia). Estudo clínico controlado e randomizado. O tempo cirúrgico e de hospitalização foram o tempo de epitelização e custos hospitalares. RESULTADOS: O fechamento das perfurações foi semelhante nos dois grupos. O tempo médio da cirurgia no grupo fáscia foi de 76,50 minutos e de 14,06 minutos no grupo com celulose bacteriana (p = 0,0001). O custo hospitalar pela tabela do SUS foi de R$ 600,00 para o grupo CB e R$ 7.778,00 para o grupo fáscia (p = 0,0001). CONCLUSÃO: A celulose bacteriana promoveu o fechamento da perfuração do tímpano, mostrando-se inovador, seguro, eficaz, efetivo, minimamente invasivo e de baixo custo.

Feasibility of bacterial cellulose membrane as biological scaffold for construction of tissue engineering corneal epithelium / 中华实验眼科杂志

Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor.The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern.Bacterial cellulose membrane has been used in medical field,but its application in tissue engineering corneal epithelium deserves more researching.Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium.Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit.Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique.The second generation ceils were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope.The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate.The ultrastructure of the cell surface was examined under the scanning electron microscope.The study was performed in accordance with the ARVO Statement.Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody.The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test,with the survival rate 100％.Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope.In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited.Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane.Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium.

Preparation and characterization of a new microporous bacterial cellulose material as a potential scaffold / 国际生物医学工程杂志

Objective When bacterial cellulose (BC) is used as a scaffold material in tissue engineering,the nano-structure of BC may not provide enough space for animal cell growth and differentiation which would not achieve a perfect application in tissue engineering.In order to solve this problem,a novel green approach is developed in this research to produce bacterial nanocellulose materials with micropores ranging 50-800 μm.Methods Several ratios of hydrogen peroxide to sodium chlorite were used to react instantly to produce a large number of bubbles in BC hydrogels,which formed micropores with diameters ranging 50-800 μm.Optical microscopy and scanning electron microscope were used to evaluate microporous BC hydrogels and verify the existence of micropores.Results The size of pores could be regulated along with the changes in the amount of reactants used in the experiment.Fourier transform infrared spectroscopy verified that no cellulose was oxidized.Water content of the microporous BC hydrogels was similar to that of the original BC hydrogels.The Young's modulus of microporous BC hydrogels was 26.1 kPa,which was lower than that of the original BC hydrogels (69.9 kPa).Thiazoyl blue tetrazolium bromide (MTT) test displayed a higher viability on the microporous BC hydrogels compared to the growth on the unmodified BC substrates.Conclusions This study provides a convenient and promising way to prepare microporous materials,which may not be limited to only BC material,but could be used in other hydrogels.The proposed approach is suitable for extensive industrialization.

Avaliação do potencial terapêutico de membranas de celulose bacteriana contendo dietilditiocarbonato (DECT)na infecção experimental por L. brasilienses / Evaluation of the therapeutic potential of diethyldithiocarbonate-containing bacterial cellulose membranes (DECT) in experimental infection by L. brasilienses

A leishmaniose continua sendo um problema de saúde pública mundial. As opções terapêuticas limitadas, a toxicidade dos fármacos disponíveis e os relatos de resistência reforçam a necessidade do desenvolvimento de novas opções terapêuticas. Neste contexto, nós demonstramos previamente que o dietilditiocarbamato (DETC), um inibidor da enzima superóxido dismutase1 (SOD1), pode diminuir a infecção por L. braziliensis, in vivo. Neste trabalho, nós testamos o DETC numa formulação tópica empregando membranas de celulose bacteriana (BC-DETC). O tratamento de macrófagos murinos infectados por Leishmania com BC-DETC resultou na morte dos parasitas intracelulares de forma direta e dose-dependente, sem evidência de efeito tóxico para as células hospedeiras. A morte parasitária, in vitro, foi associada com a diminuição da atividade da SOD1, em paralelo com o aumento da produção de superóxido e decitocinas pró-inflamatórias. A eficácia de BC-DETC, in vivo, foi demonstrada em camundongos BALB/c infectados com L. braziliensis. A aplicação tópica de BC-DETC à lesão cutânea diminuiu significativamente a úlcera na orelha e a carga parasitária no sítio de infecção. Adicionalmente, a resposta inflamatória, avaliada pela expressão de IFN-γ e TNF-α, foi suprimida in situ, bem como na resposta de memória (recall) usando células do linfonodo drenante. Finalmente, BC-DETC foi capaz de reduzir a carga parasitária em macrófagos humanos, um efeito que foi revertido na presença de antioxidante. Conjuntamente, estes resultados apontam para a viabilidade do uso de BC-DETC como uma nova formulação para a quimioterapia da leishmaniose cutânea causada por L. braziliensis.

Leishmaniasis remains a worldwide public health problem. The limited therapeutic options, drug toxicity and reports of resistance reinforce the need for the development of new treatment options. Among these options, we previously showed that diethyldithiocarbamate (DETC), a superoxide dismutase 1 inhibitor (SOD1), can decrease L. braziliensis infection, in vivo. Herein, we tested DETC in a topical formulation employing bacterial cellulose membranes (BC-DETC). Treatment of leishmania-infected murine macrophages with BC-DETC resulted in a direct and dose-dependent killing of intracellular parasites, without pronounced toxic effects to host cells. In vitro parasite killing was associated with decreased SOD1 activity paralleled by increased superoxide and pro inflammatory cytokine production. In vivo efficacy of BC-DETC was then demonstrated in L. braziliensis-infected BALB/c mice. Topical application of BC-DETC to dermal lesions significantly decreased ear thickness and parasite load at the infection site. Additionally, the inflammatory response, namely expression of IFN-γ and TNF-α, was down modulated in situ as well as in recall responses employing draining lymph node cells. Finally, BC-DETC was also capable of decreasing parasite load within human macrophages, an effect that was reversed in the presence of anti-oxidants. Collectively, these results point to the feasibility of using BC-DETC as a new topical formulation for the chemoprophylaxis of cutaneous leishmaniasis caused by L. braziliensis.

In vitro biocompatibility of bacterial cellulose and schwann cells / 第二军医大学学报

Objective To observe the biocompatibility of bacterial cellulose and Schwann cells during in vitro co-culture, so as to lay a foundation for preparing tissue-engineered nerve with bacterial cellulose. Methods The bacterial cellulose was co-cultured with the dorsal root ganglion. The morphology, proliferation, and migration of the Schwann cells were examined by scanning electron microscope and optical microscope at day 1, 2, and 5, and the protein expression of S-100 was detected by immunocytochemical method. Schwann cells were also cultivated in the leach liquor of bacterial cellulose, and then optical microscope and immunocytochemical were used to observe the morphology and S-100 protein expression after 24, 48 and 72 hours; MTT and flow cytometry were used to evaluate the biocompatibility of the bacterial cellulose and Schwann cells. Results Schwann cells could grow and migrate on the surface of bacterial cellulose's membrane. The Schwann cell growth, proliferation and S-100 expression were not significantly affected by the leach liquor of bacterial cellulose as compared with the control group. Conclusion Schwann cells can grow well on the surface of the bacterial cellulose membrane and in its leach liquor, suggesting good biocompatibility with bacterial cellulose in vitro.

The effect of bacterial cellulose membrane compared with collagen membrane on guided bone regeneration

PURPOSE: This study was to evaluate the effects of bacterial cellulose (BC) membranes as a barrier membrane on guided bone regeneration (GBR) in comparison with those of the resorbable collagen membranes. MATERIALS AND METHODS: BC membranes were fabricated using biomimetic technology. Surface properties were analyzed, Mechanical properties were measured, in vitro cell proliferation test were performed with NIH3T3 cells and in vivo study were performed with rat calvarial defect and histomorphometric analysis was done. The Mann-Whitney U test and the Wilcoxon signed rank test was used (alpha<.05). RESULTS: BC membrane showed significantly higher mechanical properties such as wet tensile strength than collagen membrane and represented a three-dimensional multilayered structure cross-linked by nano-fibers with 60 % porosity. In vitro study, cell adhesion and proliferation were observed on BC membrane. However, morphology of the cells was found to be less differentiated, and the cell proliferation rate was lower than those of the cells on collagen membrane. In vivo study, the grafted BC membrane did not induce inflammatory response, and maintained adequate space for bone regeneration. An amount of new bone formation in defect region loaded with BC membrane was significantly similar to that of collagen membrane application. CONCLUSION: BC membrane has potential to be used as a barrier membrane, and efficacy of the membrane on GBR is comparable to that of collagen membrane.

Efecto de la melaza de caña tratada con ácido sulfúrico en la producción de celulosa por Gluconacetobacter xylinus IFO 13693 / Effect of molasses cane treated with sulfuric acid in the production of cellulose by Gluconacetobacter xylinus IFO 13693 / Efeito do melaço de cana tratada com ácido sulfúrico na produção de celulose por Gluconacetobacter xylinus IFO 13693

En este estudio se evaluó el efecto de la melaza tratada con ácido sulfúrico (MZA-TR) y de las condiciones del cultivo (estático) sobre la síntesis de celulosa por Gluconacetobacter xylinus IFO 13693, para ello, se usó un reactor con 0,2 litros de medio de cultivo, con concentraciones iniciales de 13,3 % y 26,6 % de MZA-TR en el medio de cultivo a pH 5,6. El volumen del inóculo fue del 10 % del volumen total del medio; el proceso se realizó a temperatura ambiente (30 °C), con tiempos de incubación de 3, 7, 14, 21 y 28 días. Además, se evaluaron distintos parámetros fisicoquímicos y mecánicos de la celulosa. El grosor de la película de celulosa presentó un máximo de 2,5 cm, siendo el mejor resultado obtenido, en comparación con anteriores reportes en la literatura. También se encontró que al usar MZA-TR en el medio de cultivo hay un incremento considerable de la producción de celulosa en estático a los 28 días de incubación. Finalmente, se observó que el consumo de glucosa y de fructosa disminuye durante la síntesis de celulosa bacteriana (CB); durante los 3 primeros días de incubación se observó el máximo descenso, lo que permite correlacionar la producción de CB con el consumo de medio. La concentración de 13,3 %, presenta los mejores resultados en los parámetros de velocidad de crecimiento microbiano, cantidad y calidad de la celulosa producida.

This study assessed the effect of molasses treated with sulfuric acid (MZA-TR) and the condition of the crop (static) on the synthesis of cellulose by Gluconacetobacter xylinus IFO 13693, using a reactor with 0,2 liters of growing medium, with initial concentrations of 13,3 % and 26,6 % of molasses treated (MZA-TR) in the culture medium at pH 5,6. The volume of the inoculum was 10 per cent of the total volume of the environment; the process was carried out at ambient temperature (30 °C), with times of incubation of 3, 7, 14, 21 and 28 days. Also, different physico-chemical and mechanical parameters of the pulp were evaluated. The thickness of the cellulose film presented a maximum of 2.5 cm, being the best result obtained, in comparison to previous reports in the literature. Furthermore, it was found that using MZA-TR in the middle considerably increased the production of cellulose in static at 28 days of incubation. Finally, it was observed that the consumption of glucose and fructose decreased during the synthesis of Bacterial Cellulose (BC); during the first 3 days of incubation it was noted the maximum decline, which allows to correlate the production of CB with the consumption of media. The concentration of 13.3 %, presents the best results in the speed parameters of microbial growth, the quantity and quality of cellulose produced.

Neste estudo foi avaliado o efeito do melaço tratado com ácido sulfúrico (MZA-TR) e das condições da cultura (estática) sobre a síntese de celulose pelo Gluconacetobacter xylinus IFO 13693. Para isso, foi utilizado um reator com 0,2 litros de meio de cultura, com concentrações iniciais de 13,3 e 26,6% de MZA-TR, e pH igual a 5,6. O volume de inoculo foi de 10% do volume total do meio, e o processo foi realizado a temperatura ambiente (30°C), com tempos de 3, 7, 14, 21 e 28 dias. Além disso, foram avaliados os distintos parâmetros físico-químicos e mecânicos da celulose. A espessura máxima que apresentou o filme de celulose foi de 2,5 cm, sendo o melhor resultado obtido, ao comparar com os reportados anteriormente na literatura. Verificou-se também que ao usar MZA-TR no meio de cultura houve um aumento considerável na produção de celulose no meio estático aos 28 dias de incubação. Finalmente foi observado que o consumo de glicose e frutose diminuiu durante a síntese de celulose bacteriana (CB). Durante os três primeiros dias de incubação verificou-se a diminuição máxima, o que permite correlacionar a produção de CB com o consumo do meio. A concentração de 13,3% apresentou os melhores resultados nos parâmetros da velocidade de crescimento microbiano, quantidade e qualidade da celulose produzida.

Progress of bacterial cellulose and optimize production in the medical field / 国际生物医学工程杂志

Bacterial cellulose,as a new material,due to its unique nano-fiber mesh structure with good purity,physical and chemical properties such as mechanical strength and water holding capacity,biocompatibility and adaptability,has been widely used in medicine,food,paper,textile,acoustical materials and other industries.In medical field,especially,it has been rapidly developed in recent years.This review discusses the current bacterial cellulose and its performance optimization products which relates to the medical applications.