Table 2.
In vitro studies on cartilage tissue engineering using stem cells and nanofibers
| Species | Cells Source | Cells Type | Biomaterials | Stimulating Factors | Results | Ref. |
|---|---|---|---|---|---|---|
| Rabbit | Bone Marrow | Mesenchymal Stem Cells (MSCs) | Poly (Vinyl Alcohol)/Poly (E-Caprolactone): PVA/PCL | TGF-B1, FGF-2, Dexamethasone, Ascorbate 2-Phosphate, ITS+1 premix, | MSCs seeded on PVA/PCL scaffolds showed the mRNA expression of collagen type II and Aggrecan after 21 days of chondrogenic differentiation | (3) |
| Goat | Bone Marrow | MSCs | Poly (Vinyl Alcohol) - methacrylate (PVA-MA) PVA-hondroitin Sulfate- methacrylate (PVA-CS-MA) | TGF-B1, Ascorbate 2-Phosphate, Dexamethasone, L-Proline, Sodium Pyruvate, ITS-Plus Premix | A higher collagen type II/type I gene expression ratio in PVA-CS-MA compared with PVA-MA fibers alone | (7) |
| Fetal Bovine | Epiphyseal Cartilage | Chondrocytes | PCL | Ascorbate 2-Phosphate, Dexamethasone, Sodium Pyruvate, Proline, ITS-Plus Premix | Chondrocytes seeded on the PCL scaffold maintained their chondrocytic phenotype by gene expressing of collagen types IIB and IX, aggrecan, and cartilage oligomeric matrix protein | (9) |
| Human | Bone Marrow | MSCs | PCL and sodium hyaluronate (HA) | TGF- B1, Bovine Serum Albumin (BSA) | Initial release of HA is sufficient in terms of directing the implanted MSCs toward a chondrogenic end, whereas a late release of TGF-B1 is preferred to foster type II and avoid type I collagen expression | (11) |
| Human | Bone Marrow | MSCs | Poly (L-lactic) acid (PLLA) | TGF- B1 | In the presence of TGF- B1, cartilage tissue developed on PLLA scaffolds had high level of Sulfated glycosaminoglycans (sGAG), Sox-9 and collagen type II | (13) |
| Human | Umbilical Cords | MSCs | Poly L-lactide-co-glycolic acid (PLGA) and PCL | TGF-B3, TGF-B1, IGF, BMP6, Ascorbate 2-Phosphate, ITS-Plus Premix, Dexamethasone, L-Proline | Level of sGAG and sulfated proteoglycans and also the ratio of collagen type II to collagen type I expression was up-regulated in differentiated MSCs on PLGA. Cells differentiated on the scaffold | (48) |
| Human | Menstrual blood | Menstrual blood-derived stem cells (MenSCs) | PCL | TGF-B3, IGF-1, Sodium Pyruvate, Ascorbate 2-Phosphate Dexamethasone, ITS+1 premix | had high level of collagen type II and also proteoglycan production compared to 2D system | (56) |
| Human | Bone Marrow | MSCs | PCL | TGF-B1, Ascorbate 2-Phosphate, Sodium Pyruvate, L-Proline, ITS-Plus Premix | Gene expression of collagen types II and IX and also the level of sGAG was up-regulated in nanofibrous system compared with control culture | (55) |
| Human | Bone Marrow | MSCs | PLGA | TGF-B3 | MSCs seeded in PLGA nanofiber scaffold in chondrogenic induced medium began to produce high level of sGAG compared to MSCs seeded in PLGA nanofibers without chondrogenic differentiations | (60) |
| Bovine | Carpometacarpal joints of the forelimbs | Chondrocytes | PLLA | TGF-B1, IGF-1, Ascorbate 2-Phosphate, Dexamethasone, Sodium Pyruvate, Proline ITS-Plus Premix | The dynamic culture condition and IGF-1/TGF-b1 treatments upregulated collagen and sGAG production in packed cell nanofiber composite cultures | (88) |
| Human | Placentas | MSCs | nano-sized calcium-deficient hydroxyapatite (nCDHA) and/or a recombinant protein containing arginine–glycine–aspartate (RGD) into the alginate gel and PLGA | TGF-B3, Ascorbate 2-Phosphate, Dexamethasone, l-proline | The amount of sGAG and collagen type II accumulated was found to be the greatest for human Placenta-derived MSCs embedded in the alginate/nCDHA/RGD gel and injected and cultivated in the PLGA scaffold | (50) |
| Bone marrow | ||||||
| Rat | Subcutaneous Fat | |||||
| Human | Bone Marrow | MSCs | PCL | TGF- B1, Ascorbate 2-Phosphate, Sodium Pyruvate, Dexamethasone, l-proline | The expression of collagen type II and aggrecan was upregulated significantly in MSCs seeded on the nanofibrous PCL scaffold | (58) |
| Human | Cartilage | Chondrocytes | Polylactic acid (PLA) microfibers and PCL nanofibers | TGF-B1, Ascorbate 2-Phosphate, ITS+1 premix, Dexamethasone | The pore sizes in the scaffolds were tailored and increased from nanometer scale in purely nanofibrous scaffolds to hundreds of micrometers in scaffolds of nanofiber-coated microfibers. Also, SEM analysis indicated that the chondrocytes adhered and spread on composite scaffolds and produced high level of extracellular matrix. | (89) |
| Porcine | Articular Cartilage | Chondrocytes | PLGA nanofiber and membrane scaffold | Ascorbate 2-Phosphate | The DNA content and normalized sGAG content of the nanofiber based scaffolds were significantly higher than those of the membrane-type scaffolds. | (90) |
| Rabbit | Bone Marrow | MSCs | Natural Nanofibrous Articular Cartilage extracellular matrix (ACECM) and PLGA composite oriented scaffold | - | Cell proliferation test showed that the number of MSCs in ACECM and composite scaffolds was noticeably higher than that in PLGA scaffold, which was coincident with results of SEM observation and cell viability staining | (91) |
| Human | Bone Marrow | MSCs | PCL Microfibers and Nanofibers | TGF-B3, Ascorbate 2-Phosphate, L-proline, Dexamethasone, ITS+1 premix | Cellular proliferation and sGAG and collagen production were enhanced on microfiber in comparison to nanofiber scaffolds, with high initial seeding densities being required for significantchondrogenic differentiation and extracellular matrix (ECM) deposition. Moreover, the collagen type II/I ratio, as a indicator of hyaline cartilage phenotype, was significantly greater for the higher seeding densities on microfibers than nanofibers and in comparison to the lower seeding densities | (92) |
| Human | Adipose Tissue | Adipose-Derived Stem Cells (ASCs) | PCL and cartilage-derived matrix (CDM) | TGF-B1, BMP-6, Dexamethasone,, Ascorbate 2-Phosphate, L-proline | Incorporation of CDM into seeded scaffolds with hASCs stimulated sGAG synthesis and collagen type 10A1 gene expression. Also, compared with single-layer scaffolds, multilayer scaffolds enhanced cell infiltration and ACAN gene expression | (93) |
| Human | UmbilicalCord | Umbilical Cord Wharton’s Jelly Stem Cells (WJSCs) | PCL/Collagen | TGF-B3, FGF-2,L-proline, ITS+1 Premix, Dexamethasone,Ascorbate 2-Phosphate, Sodium Pyruvate | Seeded scaffolds with WJSCs and MSCs showed positive staining in 21 days for the chondrogen related proteins collagen type II and SOX9 and also sGAG values compared to controls | (94) |
| Bone Marrow | MSCs | |||||
| Rat | Bone Marrow | MSCs | PCL nanofibers encapsulated with Hyaluronic acid (HYA) and CS | - | Collagen type II was expressed more in the scaffolds with nanofibers inclusive of CS and HYA than in the scaffolds with vertically oriented nanofibers | (95) |
| Human | Bone Marrow | MSCs | PLLA Microfibers and Nanofibers | TGF-B3, ITS +1premix Dexamethasone, Ascorbic acid-2-phosphate, Sodium Pyruvate, L-proline | Chondrogenic markers of aggrecan, chondroadherin, sox9, and collagen type II were the highest for cells on micron-sized fibers in comparison to cells on nano-sized fibers | (96) |
| - | - | C3H10T1/2 murine embryonic mesenchymal progenitor cells | core-shell poly(ether sulfone)- PCL (PES-PCL) | rhBMP-2 | Results from chondrogenic differentiation of cells on scaffolds indicated that the lower modulus PCL fibers provided more appropriate microenvironments for chondrogenesis, by upregulation of Sox9, collagen type II and aggrecan gene expression and sGAG production compared to core-shell PES-PCL fibers | (97) |
| Human | Bone Marrow | MSCs | PLLA | TGF-B1, IGF-1, Dexamethasone, Ascorbic acid-2-phosphate, Sodium Pyruvate, L-proline, ITS+1 premix | The mRNA levels of aggrecan and collagen type II in TGF-B1/IGF-I treated cultures were notably higher than those treated only with TGF-B1, although these differences were not statistically significant. However, collagen type II/collagen type I ratio was high in TGFB1/IGF-I treated cultures. Also, in tow conditions, both sGAG and hydroxyproline accumulation showed significant changes over culture time | (98) |
| Human | Bone Marrow | MSCs | PCL nanofibers | TGF-B1, Ascorbic acid-2-phosphate, Dexamethasone, Sodium Pyruvate, ITS+1 premix | Constructs cultured in the presence of chondrogenic medium supplemented with TGF-B1 revealed significantly upregulated expression of aggrecan and Collagen type II and also abundant proteoglycan-rich ECM compared to constructs cultured in the presence of chondrogenic medium alone | (99) |
| Human | Articular Cartilage | Chondrocytes | Micro and Nanofibers PLLA | TGFB1, ITS+1 premix, Dexamethasone, Ascorbic acid-2-phosphate | In both types, scaffolds indicated an increase in sGAG production and Collagen type II expression over time | (100) |
| Canine | Articular Cartilage | Chondrocytes | Electrospun poly(D,L-lactide)/poly(L lactide) (PDLA/PLLA) or poly(D,L lactide)/polycaprolacton e (PDLA/PCL) with chitosan-based hydrogel | Ascorbic acid-2-phosphate | Primary canine chondrocytes produced collagen type II and proteoglycans while being cultured on scaffolds composed of electrospun PDLA/PCL and chitosan hydrogel | (101) |
| Rabbit | Articular Cartilage | Chondrocytes | PLLA nanofibers modified with cationized gelatin (CG) (CG-PLLA) | - | In vitro studies indicated that CG-PLLA could enhance viability, proliferation and differentiation of rabbit articular Chondrocytes compared with pristine PLLA nanofibers. In addition, these cell–scaffold constructs were able to maintain the expression of characteristic markers (collagen II, aggregan and SOX 9) of chondrocytes | (102) |
| Human | Bone Marrow | MSCs | PLLA nanofibers | TGF-B1, Ascorbic acid-2-phosphate, L-proline, Dexamethasone, Sodium Pyruvate, ITS+1 premix | PLLA-scaffold seeded with MSCs transfected with SOX-9 showed an increase in aggrecan mRNA expression over controls | (103) |
| Human | Bone Marrow | MSCs | PLGA nanofibers | Chondrogenic induction medium (CM, hMSC Differentiation BulletKit-chondrogenic, Lonza), TGF-B3 | Production of proteoglycan and type-II collagen and also the high expression levels of SOX9 and COL10A1 were observed in differentiated BMMSCs on nanofibers in comparison to two-dimensionally cultured cells | (61) |
| - | - | ATDC5 chondrogenic cell line | Collagen-PLA, Collagen-PLGA | - | The addition of collagen has a dual influence of making the scaffolds more hydrophilic and reinforcing the mechanical properties. Furthermore, the soft scaffolds composed of the highly biodegradable PLGA50:50 and collagen, in two ratios (40:60 and 60:40), were optimal for chondrogenesis with ECM production and enhanced cartilage specific gene expression | (62) |
| Human | Articular Cartilage | Chondrocytes | poly(3hydroxybutyrate)/poly (3hydroxyoctanoate) P(3HB)/P(3HO) | - | The finding revealed that two ratios of P(3HB)/P(3HO) enhanced the aggregation of hyaline-like cartilage matrix and type II collagen after three weeks of culture with chondrocytes | (63) |
| Rabbit | Articular Cartilage | Chondrocytes | PLLA/ silk fibroin (PLLA/SF) | - | The PLLA/SF composite scaffold supports adhesion, proliferation, and growth of chondrocyte higher than PLLA scaffold without SF | (64) |