scholarly journals Role of Hypoxia on Growth and Differentiation of Human Adipose Derived Stem Cells Grown on Silk Fibroin Scaffold Induced by Platelet Rich Plasma

2021 ◽  
Vol 53 (3) ◽  
pp. 415-427
Author(s):  
Anggraini Barlian ◽  
Marselina Irasonia Tan ◽  
Ergha Widya Sarjana ◽  
Noviana Vanawati
Keyword(s):  
Stem Cells ◽  
Silk Fibroin ◽  
Platelet Rich Plasma ◽  
Adipose Derived Stem Cells ◽  
Low Oxygen ◽  
Silk Fibroin Scaffold ◽  
Hypoxic Conditions ◽  
Proliferation And Differentiation ◽  
The Difference ◽  
Low Oxygen Concentration

Previous research has proven that 10% platelet-rich plasma (PRP) can enhance growth and differentiation of human adipose derived stem cells (hADSC) grown on silk fibroin scaffold into chondrocytes. A low oxygen concentration (hypoxia) condition is an important factor that potentially affects the ability of hADSC to grow and differentiate. The objective of this research was to analyze the difference in growth and differentiation capacity of hADSC grown on salt leached silk fibroin scaffold supplemented by 10% PRP under normoxic and hypoxic conditions. The growth capacity of the hADSC was determined by MTT assay and differentiation was tested using glycosaminoglycan (GAG) content analysis, while chondrocyte markers were visualized with the immunocytochemistry (ICC) method. This research observed hADSC proliferation under normoxic and hypoxic conditions for 21 days. Visualization of type 2 collagen showed that it was more abundant under hypoxia compared to normoxia.  HIF-1α was only detected in the hADSC cultured in hypoxic conditions. In conclusion, culture under hypoxic conditions increases the capacity of hADSC to grow and differentiate into chondrocytes. This is the first study that has shown that hypoxia is able to enhance the proliferation and differentiation of hADSC grown on 3D salt leached silk fibroin scaffold supplemented by 10% PRP.

scholarly journals KONDROGENESIS ADIPOSE-DERIVED STEM CELLS MENGGUNAKAN PLATELET-RICH PLASMA PADA SCAFFOLD SUTRA

2020 ◽  
Vol 13 (1) ◽  
pp. 31-38
Author(s):  
Imam Rosadi ◽  
Karina Karina ◽  
Komang A. Wahyuningsih ◽  
Iis Rosliana ◽  
Tias Widyastuti ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bombyx Mori ◽  
Silk Fibroin ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Platelet Rich Plasma ◽  
Adipose Derived Stem Cells ◽  
Silk Fibroin Scaffold ◽  
Salt Leaching ◽  
Fetal Bovine

AbstrakStudi tentang kemampuan adipose-derived stem cells (ADSCs) sebagai sel punca yang dapat berdiferensiasi menjadi kondrosit menggunakan platelet-rich plasma (PRP) sebagai subtitusi fetal bovine serum (FBS) telah banyak dilaporkan. Penggunaan medium pertumbuhan dengan kombinasi ADSCs, PRP dan scaffold sutra masih belum banyak dipelajari dalam rekayasa jaringan kartilago. Studi ini bertujuan untuk mempelajari pengaruh medium yang mengandung 5%, 10% dan 20% PRP terhadap proses kondrogenesis ADSCs manusia yang dikultur pada scaffold sutra Bombyx mori Indonesia. Metode penelitian diawali dengan pembuatan scaffold sutra menggunakan metode salt-leaching, isolasi dan kultur ADSCs manusia dari jaringan lemak, uji pertumbuhan ADSCs pada scaffold sutra dengan variasi konsentrasi PRP pada medium serta analisis kadar glikosaminoglikan (GAG). Hasil penelitian menunjukkan bahwa ADSCs yang dikultur menggunakan PRP lebih tinggi laju pertumbuhannya dibandingkan dikultur menggunakan FBS selama 7 hari pengamatan. Kadar GAG yang disekresikan ADSCs kelompok PRP juga lebih tinggi dibandingkan kelompok FBS. Kadar GAG tertinggi pada hari ke-21 pengamatan adalah medium yang mengandung 20% PRP kemudian 10% dan 5%, sedangkan kadar GAG kelompok kontrol cenderung stabil pada kadar yang rendah. Berdasarkan hasil tersebut, medium yang mengandung PRP memiliki potensi dalam menginduksi kondrogenesis ADSCs yang dikultur pada scaffold sutra.Abstract The studies on adipose-derived stem cells (ADSCs) differentiation into chondrocytes using platelet-rich plasma (PRP) as a substitute for fetal bovine serum (FBS) have been reported. However, the combination of ADSCs, PRP and silk fibroin scaffold has not been widely studied for developing cartilage engineering. Therefore, this research aims to study the effect of medium containing 5%, 10% and 20% PRP towards chondrogenesis of human ADSCs cultured on silk fibroin scaffold from Indonesia Bombyx mori. At first, the silk fibroin scaffold was fabricated using a salt-leaching method, then ADSCs were isolated and cultured from adipose tissues. The assays of growth curve and biocompatibility of silk fibroin scaffold toward ADSCs supplemented by PRP as well as glycosaminoglycans (GAG) concentration were conducted later. The results showed that higher absorbance of proliferation rate was on ADSCs supplemented by various PRP concentrations compare to FBS control group for seven days of observation. Level of GAG, which secreted by ADSCs supplemented by a various concentration of PRP, was also higher than the FBS group. The highest level of GAG on day 21 was observed in 20% PRP group then 10% and 5% PRP, while a group of GAG level is stable at low levels. This study concludes that PRP has the potential to induce chondrogenesis ADSCs which cultured on silk fibroin scaffold.

scholarly journals In vitro study of cartilage tissue engineering using human adipose-derived stem cells induced by platelet-rich plasma and cultured on silk fibroin scaffold

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Imam Rosadi ◽  
Karina Karina ◽  
Iis Rosliana ◽  
Siti Sobariah ◽  
Irsyah Afini ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mrna Expression ◽  
Silk Fibroin ◽  
Platelet Rich Plasma ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Silk Fibroin Scaffold

Abstract Background Cartilage tissue engineering is a promising technique for repairing cartilage defect. Due to the limitation of cell number and proliferation, mesenchymal stem cells (MSCs) have been developed as a substitute to chondrocytes as a cartilage cell-source. This study aimed to develop cartilage tissue from human adipose-derived stem cells (ADSCs) cultured on a Bombyx mori silk fibroin scaffold and supplemented with 10% platelet-rich plasma (PRP). Methods Human ADSCs and PRP were characterized. A silk fibroin scaffold with 500 μm pore size was fabricated through salt leaching. ADSCs were then cultured on the scaffold (ADSC-SS) and supplemented with 10% PRP for 21 days to examine cell proliferation, chondrogenesis, osteogenesis, and surface marker expression. The messenger ribonucleic acid (mRNA) expression of type 2 collagen, aggrecan, and type 1 collagen was analysed. The presence of type 2 collagen confirming chondrogenesis was validated using immunocytochemistry. The negative and positive controls were ADSC-SS supplemented with 10% foetal bovine serum (FBS) and ADSC-SS supplemented with commercial chondrogenesis medium, respectively. Results Cells isolated from adipose tissue were characterized as ADSCs. Proliferation of the ADSC-SS PRP was significantly increased (p < 0.05) compared to that of controls. Chondrogenesis was observed in ADSC-SS PRP and was confirmed through the increase in glycosaminoglycans (GAG) and transforming growth factor-β1 (TGF-β1) secretion, the absence of mineral deposition, and increased surface marker proteins on chondrogenic progenitors. The mRNA expression of type 2 collagen in ADSC-SS PRP was significantly increased (p < 0.05) compared to that in the negative control on days 7 and 21; however, aggrecan was significantly increased on day 14 compared to the controls. ADSC-SS PRP showed stable mRNA expression of type 1 collagen up to 14 days and it was significantly decreased on day 21. Confocal analysis showed the presence of type 2 collagen in the ADSC-SS PRP and positive control groups, with high distribution outside the cells forming the extracellular matrix (ECM) on day 21. Conclusion Our study showed that ADSC-SS with supplemented 10% PRP medium can effectively support chondrogenesis of ADSCs in vitro and promising for further development as an alternative for cartilage tissue engineering in vivo.

Integrated Trilayered Silk Fibroin Scaffold for Osteochondral Differentiation of Adipose-Derived Stem Cells

2014 ◽  
Vol 6 (19) ◽  
pp. 16696-16705 ◽  
Author(s):  
Xiaoming Ding ◽  
Meifeng Zhu ◽  
Baoshan Xu ◽  
Jiamin Zhang ◽  
Yanhong Zhao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Silk Fibroin ◽  
Adipose Derived Stem Cells ◽  
Silk Fibroin Scaffold

scholarly journals Cell penetration and chondrogenic differentiation of human adipose derived stem cells on 3D scaffold

2021 ◽  
pp. FSO734
Author(s):  
Rizka Musdalifah Amsar ◽  
Anggraini Barlian ◽  
Hermawan Judawisastra ◽  
Untung Ari Wibowo ◽  
Karina Karina
Keyword(s):  
Ascorbic Acid ◽  
Silk Fibroin ◽  
Platelet Rich Plasma ◽  
Type Ii Collagen ◽  
Adipose Derived Stem Cells ◽  
Cell Penetration ◽  
3D Scaffold ◽  
Reverse Transcription Quantitative Pcr ◽  
Proliferation And Differentiation

The ability of cells to penetrate the scaffold and differentiate into chondrocyte is important in cartilage engineering. The aim of this research was to evaluate the use of silk fibroin 3D scaffold in facilitating the growth of stem cell and to study the role of L-ascorbic acid and platelet rich plasma (PRP) in proliferation and differentiation genes. Cell penetration and type II collagen content in the silk fibroin scaffold was analyzed by confocal microscopy. Relative expressions of CDH2, CCND1, CTNNB1 and COL2A1 were analyzed by reverse transcription-quantitative PCR (RT-qPCR). The silk fibroin 3D scaffold could facilitate cell penetration. L-ascorbic acid and PRP increased the expression of CDH2 and COL2A1 on the 21st day of treatment while PRP inhibited CTNNB1 and CCND1.

Impact of low oxygen tension on stemness, proliferation and differentiation potential of human adipose-derived stem cells

2014 ◽  
Vol 448 (2) ◽  
pp. 218-224 ◽  
Author(s):  
Jane Ru Choi ◽  
Belinda Pingguan-Murphy ◽  
Wan Abu Bakar Wan Abas ◽  
Mat Adenan Noor Azmi ◽  
Siti Zawiah Omar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Oxygen Tension ◽  
Adipose Derived Stem Cells ◽  
Low Oxygen ◽  
Differentiation Potential ◽  
Proliferation And Differentiation ◽  
Low Oxygen Tension
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