In vitro chondrogenic differentiation of human adipose-derived stem                     cells with silk scaffolds

Kim, Hyeon Joo; Park, Sang-Hyug; Durham, Jennah L.; Gimble, Jeffrey M.; Kaplan, David L.; Dragoo, Jason L.

doi:10.1177/2041731412466405

Cited by 24 publications

(22 citation statements)

References 57 publications

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“…Spreafico et al (2009) showed that PRP addition in culture 386 medium was proved to increase the proteoglycan production, although the mechanism is still 387 unknown. The result of our research was supported by Kim et al (2012) who observed ADSC 388 differentiation on silk fibroin porous scaffold and showed that ADSC could grow and produce 389 chondrogenic GAG. In comparison to Kim et al (2012), we also evaluated the PRP addition and 390 found that chondrogenic differentiation showed the best when the cells were induced with PRP.…”

Section: Discussionsupporting

confidence: 63%

Peer Review #1 of "Chondrogenic differentiation of adipose-derived mesenchymal stem cells induced by L-ascorbic acid and platelet rich plasma on silk fibroin scaffold (v0.1)"

Khan¹

2018

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Articular cartilage is an avascular tissue with limited regenerative property. Therefore, a defect or trauma in articular cartilage due to disease or accident can lead to progressive tissue deterioration. Cartilage tissue engineering, by replacing defective cartilage tissue, is a method for repairing such a problem. In this research, three main aspects--cell, biomaterial scaffold, and bioactive factors--that support tissue engineering study were optimized. Adipose-derived mesenchymal stem cells (ADSC) that become cartilage were grown in an optimized growth medium supplemented with either platelet rich plasma (PRP) or L-ascorbic acid (LAA). As the characterization result, the ADSC used in this experiment could be classified as Mesenchymal Stem Cell (MSC) based on multipotency analysis and cell surface marker analysis. The biomaterial scaffold was fabricated from the Bombyx morii cocoon using silk fibroin by salt leaching method and was engineered to form different sizes of pores to provide optimized support for cell adhesion and growth.Biocompatibility and cytotoxicity evaluation was done using MTT assay to optimize silk fibroin concentration and pore size. Characterized ADSC were grown on the optimized scaffold.LAA and PRP were chosen as bioactive factors to induce ADSC differentiation to become chondrocytes. The concentration optimization of LAA and PRP was analyzed by cell proliferation using MTT assay and chondrogenic differentiation by measuring glycosaminoglycan (GAG) using Alcian Blue at 605nm wavelength. The optimum silk fibroin concentration, pore size, LAA concentration, and PRP concentration were used to grow and differentiate characterized ADSC for 7, 14, and 21 days. The cell morphology on the scaffold was analyzed using a scanning electron microscope (SEM). The result showed that the ADSC could adhere on plastic, express specific cell surface markers (CD73, CD90, dan CD105), and could be differentiated into three types of mature cells. The silk fibroin scaffold made from 12% w/v concentration formed a 500µm pore diameter (SEM analysis), PeerJ reviewing PDF | Manuscript to be reviewed and was shown by MTT assay to be biocompatible and to facilitate cell growth. The optimum concentrations of the bioactive factors LAA and PRP were 50µg/mL and 10%, respectively. GAG analysis with Alcian Blue staining suggested that PRP induction medium and LAA induction medium on 12%w/v scaffold could effectively promote not only cell adhesion and cell proliferation but also chondrogenic differentiation of ADSC within 21 days of culture. Therefore, this study provides a new approach to articular tissue engineering with a combination of ADSC as cell source, LAA and PRP as bioactive factors, and silk fibroin as a biocompatible and biodegradable scaffold.PeerJ reviewing PDF | ABSTRACT 12 Articular cartilage is an avascular tissue with limited regenerative property. Therefore, a 13 defect or trauma in articular cartilage due to disease or accident can lead to progressive tissue 14 deterioration. Cartilage tissue engineering,...

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Section: Discussionsupporting

confidence: 63%

Peer Review #1 of "Chondrogenic differentiation of adipose-derived mesenchymal stem cells induced by L-ascorbic acid and platelet rich plasma on silk fibroin scaffold (v0.1)"

Khan¹

2018

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“…Figure 5 and 6 showed the typical cell attachment and growth behaviors on different silk scaffolds. Considering that the excellent cell-compatibility of silk-based scaffolds generally results in over-rapid growth of cells [54–56], lower initial cell seeding density was preferred, which resulted in lower cell growth within 6 days. Although the rBMSCs grew well on all the scaffolds from day 1 to day 12, cells formed continuous monolayer on the silk nanofiber-based scaffolds while distributed sparsely on salt-leached scaffolds at day 12 (Figure 6C), indicating the favorable action of nanofiber in cell proliferation.…”

Section: Resultsmentioning

confidence: 99%

A mild process to design silk scaffolds with reduced β-sheet structure and various topographies at the nanometer scale

Pei

Liu

et al. 2015

Acta Biomaterialia

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Three-dimensional (3D) porous silk scaffolds with good biocompatibility and minimal immunogenicity, have promising applications in different tissue regenerations. However, a challenge remains to effectively fabricate their microstructures and mechanical properties to satisfy specific requirements of different tissues. In this study, silk scaffolds were fabricated to form extracellular matrix (ECM) mimetic nanofibrous architecture in a mild process. A slowly increasing concentration process was applied to regulate silk self-assembly into nanofibers in aqueous solution. Then glycerol was blended with the nanofiber solution and induced silk crystallization in lyophilization process, endowing freeze-dried scaffolds water-stability. The glycerol was leached from the scaffolds, leaving similar porous structure at a micrometer scale but different topographies at nanoscale. Compared to previous salt-leached and methanol annealed scaffolds, the present scaffolds showed lower β-sheet content, softer mechanical property, and improved cell growth and differentiation behaviors, implying their promising future as platforms for controlling stem cell fate and soft tissue regeneration.

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“…AdSCs is currently attracted great interest in the cartilage regeneration since it can differentiate into chondrogenic lineage [23]. The chondrogenic differentiation of ADSCs was observed on the salt leached silk fibroin scaffold, which prepared using traditional dissolution [24]. The scaffolds showed an ability to provide a suitable environment for human adipose-derived stem cells' survival and chondrogenesis.…”

Section: ) Contact Angle and Water Uptakesmentioning

confidence: 99%

Development of Salt Leached Silk Fibroin Scaffold using Direct Dissolution Techniques for Cartilage Tissue Engineering

Wibowo¹,

Judawisastra²,

Barlian³

et al. 2019

Int. J. Adv. Sci. Eng. Inf. Technol.

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Autologous transplantations, the gold standard, did not meet sufficient health tissue coverage area for cartilage damage treatments. The field of tissue engineering offers a promising alternative to fulfill this limitation by growing patient own cells on biomaterials through tissue culture, reconstructed into new cartilage tissue, and the implanted to the injury area. To support tissue regeneration, biocompatible, biodegradable, and high strength silk fibroin (SF) was proposed in this study as scaffold materials. In this research, direct dissolution in CaCl 2 /formic acid, a faster and simpler process than traditional dissolution techniques, combined with salt leaching technique. SF contents on the scaffold were varied from 2 w/v% to 12 w/v% and NaCl size as porogen was fixed in diameter of 250±58 µm. Evaluation of the SF scaffold's morphology, hydrophilicity, biodegradability, and biocompatibility were conducted. The results showed porous silk fibroin scaffold had been successfully developed. The SF scaffolds have pore size 261-293 µm with highly interconnected pores. FTIR and XRD analysis of the scaffolds showed the characteristics of silk fibroin, which reveals the α-helix amorphous and β-sheet crystalline structure and comparable to the silk fibers. The scaffold showed good hydrophilicity and high water uptakes, which essential properties for cell survival. The scaffold degraded under Protease XIV, indicate biodegradable properties. Observation of cell attachment confirms the scaffold has good biocompatibility to adipose-derived stem cells and are suitable to be used in cartilage tissue engineering.

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In vitro chondrogenic differentiation of human adipose-derived stem cells with silk scaffolds

Cited by 24 publications

References 57 publications

Peer Review #1 of "Chondrogenic differentiation of adipose-derived mesenchymal stem cells induced by L-ascorbic acid and platelet rich plasma on silk fibroin scaffold (v0.1)"

Peer Review #1 of "Chondrogenic differentiation of adipose-derived mesenchymal stem cells induced by L-ascorbic acid and platelet rich plasma on silk fibroin scaffold (v0.1)"

A mild process to design silk scaffolds with reduced β-sheet structure and various topographies at the nanometer scale

Development of Salt Leached Silk Fibroin Scaffold using Direct Dissolution Techniques for Cartilage Tissue Engineering

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