Manal Farea1, Adam Husein2, Ahmad Sukari Halim3, Nurul Asma Abdullah4, Khairani Idah Mokhtar4, Chin Keong Lim5, Zurairah Berahim6, Kasmawati Mokhtar6. 1. Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. Electronic address: ghorafi44@yahoo.com. 2. Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. Electronic address: adamkck@usm.my. 3. Reconstructive Sciences Unit, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. Electronic address: ashalim@usm.my. 4. Oral Biology Unit, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. 5. Skin Research and Innovation Centre, Research and Development Division, WIPRO Manufacturing Services Sdn Bhd, Persiaran Subang Permai, Taman Perindustrian Subang, 47610 Subang Jaya, Selangor, Malaysia. 6. Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
Abstract
OBJECTIVE: Multipotent stem cells derived from human exfoliated deciduous teeth (SHED) represent a promising cell source for tissue regeneration. In the present study we decided to test the inductive effect of chitosan and transforming growth factor-β1 (TGFβ1) as a scaffold/factor combination on SHED proliferation and osteogenic differentiation. DESIGN: Cell proliferation was quantitatively assessed by PrestoBlue, live/dead assay was performed and cell attachment to chitosan scaffold was examined by scanning electron microscopy (SEM). For osteogenic differentiation analysis, alkaline phosphatase activity was quantified, cells were stained with Alizarin Red, and the lineage specific genes/proteins ALP, COL I, BSP, and OCN were analysed by real-time PCR and Western blot. RESULTS: SHED remained viable and attached well to the chitosan structure. Moreover, TGFβ1 significantly enhanced the proliferative activity of SHED on the chitosan scaffold. Our data further revealed that chitosan and TGFβ1 enhanced the osteogenic differentiation of SHED, as evidenced by high ALP activity, strong mineral deposition, and the up-regulation of ALP, COL I, BSP, and OCN gene/protein expression. CONCLUSION: Together, data from our study indicate that the combination of chitosan scaffolds and TGFβ1 enhanced proliferation and osteogenic differentiation of SHED. These findings suggest that the combined application of chitosan scaffold and TGFβ1 in conjunction with SHED might be beneficial for in vivo bone regeneration.
OBJECTIVE: Multipotent stem cells derived from human exfoliated deciduous teeth (SHED) represent a promising cell source for tissue regeneration. In the present study we decided to test the inductive effect of chitosan and transforming growth factor-β1 (TGFβ1) as a scaffold/factor combination on SHED proliferation and osteogenic differentiation. DESIGN: Cell proliferation was quantitatively assessed by PrestoBlue, live/dead assay was performed and cell attachment to chitosan scaffold was examined by scanning electron microscopy (SEM). For osteogenic differentiation analysis, alkaline phosphatase activity was quantified, cells were stained with Alizarin Red, and the lineage specific genes/proteins ALP, COL I, BSP, and OCN were analysed by real-time PCR and Western blot. RESULTS: SHED remained viable and attached well to the chitosan structure. Moreover, TGFβ1 significantly enhanced the proliferative activity of SHED on the chitosan scaffold. Our data further revealed that chitosan and TGFβ1 enhanced the osteogenic differentiation of SHED, as evidenced by high ALP activity, strong mineral deposition, and the up-regulation of ALP, COL I, BSP, and OCN gene/protein expression. CONCLUSION: Together, data from our study indicate that the combination of chitosan scaffolds and TGFβ1 enhanced proliferation and osteogenic differentiation of SHED. These findings suggest that the combined application of chitosan scaffold and TGFβ1 in conjunction with SHED might be beneficial for in vivo bone regeneration.
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