The cartilaginous tissue formation using sry (Sex Determining Region Y)-BOX9 and telomerase reverse transcriptase genes transfected chondrocytes: in vivo approach
The shortage of organ supply reduces the success rate of organ transplantation. Hence, tissue regeneration has been initiated with the intention of improving the available treatment modalities. Articular cartilage is a suitable tissue for this purpose due to its limited self-heal ability. This stu...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Penerbit Universiti Kebangsaan Malaysia
2020
|
| Online Access: | http://journalarticle.ukm.my/15397/ http://journalarticle.ukm.my/15397/1/11.pdf |
| Summary: | The shortage of organ supply reduces the success rate of organ transplantation. Hence, tissue regeneration has been
initiated with the intention of improving the available treatment modalities. Articular cartilage is a suitable tissue for this
purpose due to its limited self-heal ability. This study aims to evaluate the cartilaginous properties of in vivo constructs
formed using chondrocytes transfected with the combination of sry (sex determining region y)-box9 (SOX9) and
telomerase reverse transcriptase (TERT) genes (SOX9/TERT-transfected chondrocytes) seeded on a three-dimensional
(3D) poly(lactic-co-glycolic) acid (PLGA)-based scaffold. The rabbit’s articular chondrocytes (n=6) were transfected
with SOX9 and TERT genes via lipofection. The non-transfected chondrocyte (NTC) was used as a control. A total of
1×105 cells were seeded on a PLGA and PLGA/fibrin hybrid scaffolds to form constructs. The resulted constructs were
SOX9/TERT-PLGA/fibrin, NTC-PLGA/fibrin, SOX9/TERT-PLGA and, NTC-PLGA. All constructs were cultured for three
weeks prior to subcutaneous implantation into the athymic mice for two and four weeks. The constructs’ structural and
functional aspects were evaluated using macroscopic observation, compression-stress analysis, histology, quantitative
sulphated glycosaminoglycan (sGAG) assay and cartilage-specific genes (ACAN, COL2A1, SOX9), TERT, and MMP13
expression analysis. The constructs demonstrated a cartilage-like appearance. The constructs’ rigidity corresponded to
the homogenous cells and extracellular matrix distribution in the week-4 constructs. Correspondingly, the cartilaginous
matrix components were visualised at the pericellular matrix region of the construct, supported by the increment of
quantitative sGAG content. The SOX9/TERT-PLGA/fibrin exhibited better genes expression and cartilaginous phenotypes
than the other construct groups. The SOX9/TERT-PLGA/fibrin construct facilitated cartilaginous tissue formation. |
|---|