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Huang S.,Chinese Institute of Basic Medical Sciences | Fu X.,Chinese Institute of Basic Medical Sciences | Fu X.,Burns Institute
Journal of Alloys and Compounds | Year: 2010

Microparticles can serve as substrates for cell amplification and deliver the cell aggregation to the site of the defect for tissue regeneration. To develop favorable microparticles for cell delivery application, we fabricated and evaluated three types of microparticles that differ in surface properties. The microparticles with varied surface morphology (smooth, pitted and multicavity) were created from chemically crosslinked gelatin particles that underwent various drying treatments. Three types of microparticles were characterized and assessed in terms of the cell behavior of human keratinocytes and fibroblasts seeded on them. The cells could attach, spread and proliferate on all types of microparticles but spread and populated more slowly on the microparticles with smooth surfaces than on those with pitted or multicavity surfaces. Microparticles with a multicavity surface demonstrated the highest cell attachment and growth rate. Furthermore, cells tested on microparticles with a multicavity surface exhibited better morphology and induced the earlier formation of extracellular-based cell-microparticle aggregation than those on microparticles with other surface morphology (smooth and pitted). Thus, microparticles with a multicavity surface show promise for attachment and proliferation of cells in tissue engineering. © 2009 Elsevier B.V. All rights reserved.

Li H.,Shantou University | Chen L.,Shantou University | Zhang M.,Shantou University | Tang S.,Shantou University | Fu X.,Burns Institute
Cell and Tissue Research | Year: 2013

Interactions between the extracellular matrix (ECM) and epithelial cells are necessary for the proper organization and function of the epithelium. In the present study, we show that human eccrine sweat gland epithelial cells cultured in matrigel, a representation of ECM components, constitute a good model for studying three-dimensional reconstruction, wound repair and regeneration and differentiation of the human eccrine sweat gland. In matrigel, epithelial cells from the human eccrine sweat gland form tubular-like structures and then the tubular-like structures coil into sphere-like shapes that structurally resemble human eccrine sweat glands in vivo. One sphere-like shape can be linked to another sphere-like shape or to a cell monolayer via tubular-like structures. Hematoxylin and eosin staining has revealed that the tubular-like structures have a single layer or stratified epithelial cells located peripherally and a lumen at the center, similar to the secretory part or duct part, respectively, of the eccrine sweat gland in sections of skin tissue. Immunohistochemical analysis of the cultures has demonstrated that the cells express CK7, CK19, epithelial membrane antigen and actin. Thus, matrigel promotes the organization and differentiation of epithelial cells from the human eccrine sweat gland into eccrine sweat gland tissues. © 2013 Springer-Verlag Berlin Heidelberg.

Objective: To compare the effect of intravenous resuscitation with pyruvated Ringer solution with lactated Ringer solution on hemodynamics and organ function during shock resuscitation in dogs with burns. Methods: 28 Beagle dogs were subjected to a 50% total body surface area (TBSA) burns and divided into three groups: burn injury without fluid resuscitation (NR, n= 8), burn with lactated Ringer solution (RL, n=10), and pyruvated Ringer solution (RP, n=10). They were given intravenous fluid resuscitation according to Parkland formula 30 minutes after burns. The hemodynamics, organ function and mortality were observed in conscious state before burn injury, and 2, 6, 8, 12, 24 hours after burn injury. Results: Within 24 hours after burns, all the dogs in the NR group died and those in RL and RP groups were all alive. At 2 hours after burn, the mean arterial pressure (MAP), cardiac index (CI), and dp/dt max of left ventricular contractility were significantly reduced in NR, RL, and RP groups compared with those before injury [MAP(mmHg, 1 mmHg=0.133 kPa): 45.33 ± 7.78 vs. 141.67 ± 5.98, 91.33 ± 10.25 vs. 142.33 ± 6.16, 98.67 ± 9.54 vs. 142.83 ± 5.47; CI (mL·s-1m -2): 8.17 ± 0.83 vs. 48.34 ± 3.33, 16.84 ± 2.17 vs. 47.34 ± 1.67, 19.00 ± 1.50 vs. 47.34 ± 1.33; dp/dt max (mmHg/s): 426.83 ± 51.91 vs. 1 372.50 ± 39.61, 594.00 ± 88.23 vs. 1 363.83 ± 44.92, 645.00 ± 66.82 vs. 1 395.83 ± 19.49, all P<0.05], and the systemic vascular resistance (SVR) and alanine transaminase (ALT), creatinine (Cr), serum MB isoenzyme of creatine kinase (CK-MB), and diamine oxidase (DAO) were significantly higher [SVR(kPa·s·L-1): 1 322.50 ± 36.37 vs. 281.45 ± 8.84, 777.50 ± 41.84 vs. 289.72 ± 6.70, 571.40 ± 40.01 vs. 286.27 ± 8.66; ALT (U/L): 89.50 ± 4.11 vs. 40.57 ± 3.63, 89.25 ± 4.88 vs. 37.92 ± 2.62, 86.30 ± 5.61 vs. 38.47 ± 3.50; Cr (μmol/L): 75.62 ± 4.61 vs. 41.58 ± 2.78, 77.00 ± 5.92 vs. 46.55 ± 3.17, 74.13 ± 2.56 vs. 45.65 ± 1.83; CK-MB (kU/L): 13.122 ± 2.821 vs. 1.557 ± 0.087, 8.885 ± 0.272 vs. 1.497 ± 0.086, 8.692 ± 0.180 vs. 1.490 ± 0.046; DAO (kU/L): 2.26 ± 0.14 vs. 0.25 ± 0.02, 1.50 ± 0.07 vs. 0.25 ± 0.01, 1.37 ± 0.07 vs. 0.25 ± 0.02, all P<0.05]. All parameters in the NR group kept on worsening till death, while hemodynamics and organ function of two intravenous resuscitation groups were gradually improved, SVR and DAO in the RP group were significantly superior to those of the RL group since 2 hours after burns, CI in the RP group was superior to that of the RL group since 6 hours after burns, and dp/dt max and CK-MB in the RP group were more significantly preserved than those in the RL group since 8 hours after burns, Cr was significantly reduced at 24 hours after burns in the RP group compared with the RL group [2-hour SVR (kPa·s·L-1): 571.40 ± 40.01 vs. 777.50 ± 41.84, 2-hour DAO(kU/L): 1.37 ± 0.07 vs. 1.50 ± 0.07, 6-hour CI (mL·s-1·m-2): 38.67 ± 2.17 vs. 33.17 ± 1.67, 8-hour dp/dt max (mmHg/s): 1 153.83 ± 45.72 vs. 1 054.80 ± 57.70, 8-hour CK-MB (kU/L): 6.387 ± 0.267 vs. 6.806 ± 0.237, 24-hour Cr (μmol/L): 53.42 ± 4.99 vs. 60.77 ± 3.11, all P<0.05]. Conclusion: The pyruvated Ringer solution was superior to the lactated Ringer solution in improving hemodynamics and organ function for intravenous resuscitation in dogs with 50%TBSA full thickness burns.

Zhang C.,Burns Institute | Chen P.,Beijing Institute of Radiation Medicine | Fei Y.,Burns Institute | Liu B.,Burns Institute | And 5 more authors.
Aging Cell | Year: 2012

Aged epidermal cells have the capacity to dedifferentiate into stem cell-like cells. However, the signals that regulate the dedifferentiation of aged epidermal cells remain unclear. Here, we provide evidence that Wnt/β-catenin is critical for aged epidermal cell dedifferentiation in vivo and in vitro. Some aged epidermal cells in human ultrathin epidermal sheets lacking basal stem cells transplanted onto wounds dedifferentiated into stem cell-like cells that were positive for CK19 and β1 integrin but negative for CK10. In addition, Wnt/β-catenin pathway was activated during this process. There was increased expression of Wnt-1, Wnt-4, Wnt-7a, β-catenin, cyclin D1, and c-myc. Secreted frizzled-related protein 1, a Wnt/β-catenin pathway inhibitor, blocked dedifferentiation in vivo. Then, the activator, a highly specific glycogen synthase kinase (GSK)-3β inhibitor, of Wnt/β-catenin pathway was added to the culture medium of aged epidermal cells. Surprisingly, we found that the activator induced higher expression of CK19, β1 integrin, Oct4, and Nanog proteins. The induced aged epidermal cells exhibited high colony-forming efficiency, long-term proliferative potential and could regenerate a skin equivalent (as do epidermal stem cells). These results suggested that activation of Wnt/β-catenin pathway induced the dedifferentiation of aged epidermal cells, which suggest a new approach to generate epidermal stem cell-like cells. © 2011 The Authors. Aging Cell © 2011 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland.

Huang S.,Chinese Institute of Basic Medical Sciences | Xu Y.,Burns Institute | Wu C.,Burns Institute | Sha D.,Burns Institute | And 2 more authors.
Biomaterials | Year: 2010

Despite the rapid development of engineered skin, such skin still lacks skin appendages. Sweat glands, one of the skin appendages, play key roles in the maintenance of homeostasis and temperature regulation. In this study, we tested whether sweat glands could be integrated into engineered skin constructs to improve the quality of tissue regeneration. Using gelatin microspheres(containing epidermal growth factor [EGF]) as multifunctional vehicles, we cultured sweat gland cells (SGCs) on them and delivered SGCs-microspheres complex (SMC) into the engineered skin construct, which was created in vitro by culturing human keratinocytes on top of a fibroblast-embedded collagen-based matrix in an organotypic co-culture model. This engineered skin construct was then transplanted onto full-thickness cutaneous wounds in an athymic murine model. EGF-loaded microspheres displayed more cellular growth-promoting efficiency, and thus SMC was an available means for SGCs delivery. Constitution of the engineered skin constructs formed a skin-like pattern in vitro. Remarkably, SMC could differentiate toward a sweat gland-like structure in vitro within the hybrid matrix. Furthermore, the degree of wound healing in mice with this skin construct implantation was better than that with controls. This engineered skin construct could be used as a promising tool for regeneration of sweat glands in skin repair and a valuable engineered strategy for constitution of appendage-containing engineered skin models. © 2010 Elsevier Ltd.

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