Repair of sciatic nerve defect in rats by adipose tissue decellularized matrix hydrogel / 中华显微外科杂志

Objective:To observe the effect of adipose tissue decellularized matrix hydrogel (DAT-gel) on the repair of sciatic nerve defect in rats.Methods:From April, 2019 to April, 2020, aseptic granular adipose tissue was collected from healthy adult women who underwent thigh or abdominal liposuction in the Department of Plastic Surgery, the First Medical Centre of the PLA General Hospital. Decellularisation and enzymatic digestion of adipose tissue were performed to prepare DAT-gel. Scanning electron microscope (SEM) was used to observe the ultrastructure of the hydrogel, and rheology was employed to test the gel dynamics and viscoelasticity of the hydrogel. A rat model of sciatic nerve defect was established and randomly divided into 3 groups: simple chitin catheter group (Chitin group), DAT-gel plus chitin catheter group (DAT-gel group) and autologous nerve reverse connection group (Autograft group) with 10 rats in each group. At the 12th week after surgery, the general view, function and morphology of the regenerated nerve were observed to evaluate the repairing status of the injured nerve. One-way analysis of variance (one-way ANOVA) was used for data analysis. If the difference between the groups was statistically significant, the Turkey method was further used for pairwise comparison. P<0.05 was considered as statistically significant. Results:The results of SEM showed that the DAT-gel had a three-dimensional structure in porous fibre network. The results of rheological test results showed that the complex viscosity of the hydrogel at 4 ℃ and 37 ℃ were 148.91 mPa·s and 801.29 mPa·s, respectively. DAT-gel underwent a sol-gel phase transition when the temperature had been increased. The results showed that DAT-gel had a good temperature-sensitive effect, and its critical point of sol-gel phase transition was similar to the internal temperature of rat. The results of animal experiments showed that the morphology and function of the regenerated nerve in the DAT-gel group were superior to Chitin group at 12 weeks after surgery, according to macroscopic view of the regenerated nerve, electrophysiology of the nerve, the morphology of the new axon and the target muscle, etc.. There was statistically significant between groups ( P<0.05). Conclusion:DAT-gel can significantly promote a repair of sciatic nerve defects in rats.

Decellularized matrix of human fatty liver used for three-dimensional culture of hepatocellular carcinoma cells / 南方医科大学学报

OBJECTIVE@#To construct a decellularized matrix of human fatty liver as the scaffold for three-dimensional (3D) culture of hepatocarcinoma cells.@*METHODS@#Human fatty liver decellularized matrix (hFLM) was prepared by repeated freezingthawing, perfusion with gradient SDS and 1% Triton X-100 through the portal vein and hepatic artery, and repeated agitation with Triton X-100. HepG2 cells were cultured in the prepared hFLM, and the cell survival, morphology, proliferation and cellular expressions of the adhesion molecules were detected.@*RESULTS@#The decellularization procedure shortened the time for scaffold preparation and preserved the 3D ultrastructure and the composition of the extracellular matrix. HepG2 cells cultured in hFLM scaffold maintained proliferation for up to 15 days and showed a growth pattern with a long lag phase and a slow growth rate, which was similar to the growth pattern . The cultured HepG2 exhibited a low expression of E-cadherin and a high expression of vimentin, which was consistent with the xenograft but opposite to 2D cultured cells. However, the lack of adequate nutrient transport in this hepatocarcinoma cell model led to a slowdown of cell proliferation in the later stage. The PCNA index of HepG2 cells cultured in hFLM was lowered by 29.3% on day 12 as compared with that on day 6.@*CONCLUSIONS@#We established a new protocol for preparing hFLM and confirmed the feasibility of constructing hepatocarcinoma cell models using the hFLM scaffold.

Evaluación del método químico-enzimático de descelularización para la obtención de matrices extracelulares de tráquea en el modelo porcino / Avaliação do método químico-enzimático de descelularização para a obtenção de matrizes extracelulares de traqueia no modelo suíno / Evaluation of the chemical-enzymatic decellularization method for obtaining extracellular trachea matrixes in a porcine model

Diferentes enfermedades afectan la tráquea y deterioran la calidad de vida. La ingeniería tisular es una alternativa terapéutica para los pacientes con esas enfermedades: matrices de tráquea descelularizadas y sembradas con células del receptor no generan respuesta inmune y pueden prevenir incluso el rechazo de zootrasplantes. Objetivo: evaluar un método de descelularización para obtener matrices extracelulares de tráquea en el modelo porcino. Materiales y métodos: a partir de 5 tráqueas porcinas se formaron dos grupos de estudio, controles y tratados con un método químico-enzimático. Se hizo análisis histológico con hematoxilina-eosina, coloración tricrómica de Masson y safranina O. Se evaluaron las propiedades biomecánicas de ambos grupos, mediante la determinación del módulo de Young, la fuerza máxima y el porcentaje de deformación. Resultados: en las muestras tratadas se observó una disminución del 66 % del contenido celular en comparación con los controles. Se preservó el colágeno y se detectó reducción de los glucosaminoglucanos. Las pruebas biomecánicas revelaron una diferencia estadísticamente significativa del porcentaje de deformación, sin alteración de los demás parámetros. Conclusiones: el método evaluado demostró ser eficiente para descelularizar tráqueas de cerdo, con una disminución importante en el costo y el tiempo de tratamiento, por lo que podría ser una buena opción en las condiciones socioeconómicas de Colombia...

Different diseases may affect the trachea and, therefore, the quality of life. Tissue engineering may be a therapeutic alternative for patientswith such diseases, using decellularized trachea matrixes seeded with cells from the recipient, which do not generate immune response and may even prevent rejection of zoo-transplants. Objective: To evaluate a decellularization method to obtain tracheal extracellular matrixes in the porcine model. Materials and methods: Two study groups, treated and control, were obtained from 5 porcine tracheas.A chemical-enzymatic method for decellularization was used. Histological analyses were performed with hematoxylin-eosin, Masson’s trichromic stain and safranin O. Biomechanical properties of both groups were evaluated by determining the Young modulus, maximum strength and deformation rate. Results: Compared to the controls, there was a 66 % decrease in the cell content in the treated specimens. Collagen was preserved and a reduction of glycosaminoglycans was detected. Biomechanical tests revealed a significant difference in the percentage of deformation, with no alteration of the remaining parameters. Conclusions: The evaluated decellularization method proved to be efficient to reduce the cellular content of porcine tracheas, with a considerable decrease in cost and production time. These advantages could make it a good option for the socio-economic Colombian conditions...

Diferentes doenças afetam a traqueia e deterioram a qualidade de vida. A engenheira tisular é uma alternativa terapêutica para os pacientes com essas doenças: matrizes de traqueia descelularizadas e semeadas com células do receptor não geram resposta imune e podem prevenir incluso a rejeição de zoo-transplantes. Objetivo: avaliar um método de descelularização para obter matrizes extracelulares de traqueia no modelo suíno. Materiais e métodos: a partir de 5 traqueias suínas se formaram dois grupos de estudo, controles e tratados com um método químico-enzimático. Se fez análise histológico com hematoxilina-eosina, corante tricrómica de Masson e safranina O. Se avaliaram as propriedades biomecânicas de ambos grupos, mediante a determinação do módulo de Young, a força máxima e a porcentagem de deformação. Resultados: Nas amostras tratadas se observou uma diminuição de 66 % do conteúdo celular em comparação com os controles. Se preservou o colágenoe se detectou redução dos glucosaminoglucanos. As provas biomecânicas revelaram uma diferença estatisticamente significativa da porcentagem de deformação, sem alteração dos demais parâmetros. Conclusões: O método avaliado demostrou ser eficiente para descelularizar traqueias de porco, com uma diminuição importante no custo e o tempo de tratamento, pelo que poderia ser uma boa opção nas condições socioeconómicas da Colômbia...