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  • ISSN: 2379-0490
    Early Online
    Volume 4, Issue 1
    Research Article
    Tetsutaro Kikuchi and Tatsuya Shimizu*
    Abstract: Purification of skeletal muscle myoblasts from other cells such as fibroblasts is essential in basic research and cell-based regenerative medicine. In this report a simple non-serological method is proposed to purify desmin-positive myoblasts using Brefeldin A.
    Ordinary passaged human skeletal muscle-derived cells were cultured for 3 days with five exocytic pathway inhibitors (Brefeldin A, Monensin, AACOCF3, Exo-1 and Golgicide A). The myoblasts identified by desmin exhibited relatively superior growth compared to the other cells, both in 7 - 15 ng/mL of Brefeldin A and in 850 – 2500 ng/mL of Golgicide A. In the case of 10 ng/mL Brefeldin A, the cell numbers increased 6.7-fold in desmin-positive cells and 2.4-fold in desmin-negative cells while they increased 11.7-fold and 15.6-fold in absence of Brefeldin A, respectively. In addition, 7-day cultures in 10 ng/mL Brefeldin A of four independent cell strains suggested that moderate starting purities resulted in high purities over 90%, while extremely low starting purities (< 1%) did not support stable purifications. Lastly, a strain purified through a 7-day culture with 10 ng/mL Brefeldin A was subjected to myotube induction culture. Fluorescent imaging using a confocal microscope revealed their ability to form multinucleated myotubes and myofibers with sarcomeric structures.
    In conclusion, the purification of desmin-positive cells by culturing human skeletal muscle-derived cells with Brefeldin A provides a simple method to obtain purified myoblasts for use in basic research and cell-based regenerative medicine.
    Short Communication
    Aline Evangelista Aguiar, Mariana de Oliveira Silva, Bianca Cheregatti Longo, Maria do Carmo Gonçalves, Celso Aparecido Bertran*
    Scaffold porosity is an essential factor for tissue growth since it allows cell proliferation and vascularization. In this work, three different scaffolds were produced by easy methods. The first one was prepared using Bioglass 45S5 particles and H2O2 as a foaming agent. In the second, glass nanoparticles were arranged as porous structure by the self-assembling of cellulose nanocrystals. In the last, a polymeric xanthan/chitosan composite scaffold was prepared by swelling method. The results showed that the scaffolds presented pore morphology, wall thickness and size distribution that are dependent on the materials and preparation method used, and that they are suitable for a range of applications.
    Review Article
    Sharma C, Dhasmana A, Gupta SK, Purohit SD, Yadav I, Singh H, and Mishra NC*
    Bone loss owing to trauma, congenital defects and sports-related injuries has become a major health quandary all over the world. Existing therapies, although somewhat successful, do not provide the optimum remedy to orthopedic disorders. Conventional treatments typically rely on donor tissues obtained either from the patient or from another source, which raises the issue of donor-cell-supply, immune rejection and disease transfer. This has incited orthopaedic surgeons to look for viable alternatives. A smart option to overcome these problems is served by a tissue-engineered bone. Bone regeneration can be attained by an appropriate combination of cells, scaffold and growth factors. To regenerate full functional bone, researchers worked for decades to find suitable combination of cells, biomaterials for scaffold fabrication, scaffold structure and growth factor: the work is still in progress. Various biomaterials including bioceramics have emerged as an effective module for fabricating scaffold for bone tissue engineering. Stem cells have gained importance as a potent cell source for bone regeneration. Stem cells along with multiple growth factor approach are applied nowadays to regenerate bone. The delivery of these growth factors in conjunction with gene therapy has come forward as an ideal approach for augmenting bone tissue. This review highlights the advances in bone tissue engineering by focusing on three key components cell sources, scaffold biomaterials as well as growth factors used in bone tissue engineering. It also reflects an array of problems and future perspectives to overcome the existing stumbling blocks in bone regeneration.
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