Aparnathi Mansi K, Patel Jagdish S
P. D. Patel Institute of Applied Science, Charotar University of Science and Technology, Changa, Gujarat, India.
J Stem Cells. 2016;11(3):111-119.
Therapeutic potential of adipose derived stem cells (ADSCs) has widely been explored for treatment of orthopedic ailments. Transplantation of cells encapsulated in a scaffold facilitates 3 dimensional modelling of the tissue for the cases where well-defined spatial distribution of cells is desired for implantation. Present study aims to encapsulate canine ADSCs (cADSCs) in biodegradable methacrylated gelatin gel (GelMA) scaffold followed by their osteogenic differentiation for fabrication of a three dimensional bone tissue construct. Different percentages (5, 10 and 20%) and different methacrylation levels of gel (GelMA and GelMA) were tested for degradation. Porosity of 10% GelMA was compared by SEM imaging. Gels with the fastest degradation rate (5% GelMA and GelMA) were chosen for cell encapsulation since degradation of scaffold is of prime importance when the gel is intended to be used for implantation. Finally, cADSCs encapsulated in 5% GelMA demonstrated best morphology and were differentiated osteogenically. We developed a modified protocol for isolation of RNA from cells encapsulated in GelMA. Osteogenic differentiation was affirmed by the presence of osteo-specific gene expression by reverse transcriptase PCR in addition to von Kossa staining of the construct. GelMA is an excellent biodegradable scaffold for encapsulation of cADSCs without altering their osteogenic potential. This osteo-induced cellular scaffold implant opens a new therapeutic horizon in the area of tissue engineering in orthopedics.
脂肪来源干细胞（ADSCs）在治疗骨科疾病方面的治疗潜力已得到广泛探索。对于那些植入时需要细胞有明确空间分布的情况，将细胞封装在支架中进行移植有助于组织的三维建模。本研究旨在将犬脂肪来源干细胞（cADSCs）封装在可生物降解的甲基丙烯酸化明胶凝胶（GelMA）支架中，然后使其进行成骨分化，以构建三维骨组织构建体。测试了不同百分比（5%、10%和20%）以及不同甲基丙烯酸化水平的凝胶（GelMA和GelMA）的降解情况。通过扫描电子显微镜成像比较了10% GelMA的孔隙率。选择降解速度最快的凝胶（5% GelMA和GelMA）进行细胞封装，因为当凝胶打算用于植入时，支架的降解至关重要。最后，封装在5% GelMA中的cADSCs表现出最佳形态并进行了成骨分化。我们开发了一种从封装在GelMA中的细胞中分离RNA的改良方案。除了对构建体进行冯·科萨染色外，通过逆转录聚合酶链反应检测到骨特异性基因表达，从而证实了成骨分化。GelMA是一种出色的可生物降解支架，用于封装cADSCs时不会改变其成骨潜力。这种骨诱导细胞支架植入物为骨科组织工程领域开辟了新的治疗前景。
