Table 1.

Summary of composite nanomaterials developed for cartilage healing and regeneration.

Scaffolds	Fillers	Tested Cell Cultures	In Vitro Tests	In Vivo Tests	Preparation	Effect	Ref.
Collagen, chitosan	Micro graphene oxide, BMP-2 (1:1)	Chondrocytes	SEM, immunofluorescence	Rats, knee, femur cartilages	3D printed	Enhanced chondrocyte proliferation	[83]
Polycaprolactone	Graphene nanoplatelets, polycarboxylate modified graphene nanoplatelets 0.5, 5, 10 wt%	Human chondrocytes knee, hip	Cytotoxicity, cell proliferation	-	Injection molding process to make filaments in form of sticks, 3D printing	Improved mechanical properties, support of proliferation of chondrocytes	[84]
Acellular cartilage extracellular matrix, distal femoral condyle of market-weight pigs	Graphene oxide 0, 1, 2, 4, 6 mg/mL	Chondrocytes	Cell viability, adhesion, and proliferation, chondrogenesis	Implantation in rats, cartilage defect model in rabbits and histological evaluation	-	Improvement of internal structure and mechanical properties	[85]
Sericin	Reduced graphene oxide in ratio 10:1, 50:1, 100:1	Mesenchymal stem cells derived from bone marrow of humans	Mesenchymal stem cells differentiation, growth, adhesion	-	-	Increased levels of collagen and glycosaminoglycan, chondrogenic differentiation stimulation	[86]
Gelatin, methacrylate polyethylene (glycol) diacrylate	Graphene oxide	Primary human bone marrow mesenchymal stem cells	Mesenchymal stem cells proliferation, chondrogenic differentiation, collagen II secretion, glycosaminoglycan synthesis, total collagen levels, RT-PCR	-	3D printed scaffolds	Favorable mechanical properties, biocompatibility, increased collagen, glycosaminoglycan, protein levels; chondrogenic differentiation of mesenchymal stem cells	[87]
Chitosan, gelatin, anionic non-sulfated glycosaminoglycan	Graphene 0, 0.024, 0.06, 1%	Bone marrow mesenchymal stem cells	-	-	Bioink, 3D-printing	Enhanced water absorption, porosity, compression modulus, cytocompatibility, cell growth, higher cells proliferation survival	[80]
Poly(ε-caprolactone)	Graphene nanopowders 1, 3, 5, 10 wt%	Mouse bone marrow mesenchymal stem cells	Cell culture studies, MTT Assay, Live/Dead® assays, glycosaminoglycan formation, cell attachment and morphology	-	Printing ink, 3D-printing, robocasting method	Highest cell viability rates of cells seeded onto composite scaffolds, cells proliferated well, attached to scaffold surfaces	[88]
Polycaprolactone	Graphene and single-wall carbon nanotubes, 0.5% and 1.0% poly-l-lysine coated	Mesenchymal stem cells	Mesenchymal stem cells cell adhesion, proliferation, and chondrogenic differentiation	-	Electrospinning, microfibrous scaffolds	Improved mechanical properties, more homogenous fiber morphology, surface properties, good cytocompatibility	[79]
α-Chitin, poly(caprolactone)	Chondroitin sulfate, transforming growth factor-β encapsulation	Adipose derived stem cell from inguinal fat pads of female New Zealand albino rabbit	Cell viability, attachment, and proliferation study, chondrogenic differentiation and analysis of a murine rheumatoid arthritis model	-	Lyophilization technique	Prolonged release of TGF-β achieved, macroporous, extremely porous structure, enhanced cell attachment, proliferation, differentiation	[81]
-	Graphene oxide granules	Umbilical cord mesenchymal stem cells	-	Male New Zealand white rabbits: expression levels of nitric oxide, interleukin-6, tumor necrosis factor-α, glycosaminoglycan, collagen-II in serum and articular fluid	Mixing	Reduction in inflammatory level, improve of level of biochemical environment in articular cavity, promotion of cartilage repair	[89]
2% chitosan	0, 0.1, 0.2, 0.3 (w/v) % suspensions of graphene oxide in deionized water	Human articular chondrocytes	MTT assay	-	Ultra-sonication process	Improvement of physical, mechanical properties, increased proliferation of human articular chondrocytes	[90]
Poly(lactide-co-glycolide acid)	Graphene oxide	Bone marrow mesenchymal stem cells	Rabbit bone marrow mesenchymal stem cells	Rabbit supraspinatus tendon repair model	Electrospining	Accelerated proliferation and osteogenic differentiation, promoted healing, increased bone and cartilage generation, improved collagen arrangement	[91]
Collagen-I, genipin	Carbon dots	Bone marrow derived stem cells	Chondrocyte differentiation medium, intracellular ROS production, Cell Counting Kit (CCK)-8 assay, cell viability	Articular cartilage intracellular ROS production	Mixing	ROS production by photodynamic therapy, enhanced cartilage regeneration, chondrogenic differentiation, increased stiffness, reduced degradation	[92]
Collagen-II, chitosan, poly(lactic-co-glycolic acid)	-	Rabbit chondrocytes labelled with magnetic Iron oxide nanoparticles, TANBead® USPIO-101 (Amine group, Taiwan Advanced Nanotech Inc., Taipei, Taiwan)	Cell proliferation assay reagent WST-1, cell viability, cytotoxicity, relative proliferation activity	New Zealand White rabbits: levels of chondrogenetic marker genes including Sox-9, aggrecan, collagen-II	Mixing	Incorporation of chondrocytes into cartilage by magnetic force	[93]
Chitosan, collagen-I	Bioactive glass nanoparticles	Human osteosarcoma cell culture (SAOS) and kidney cells line of human embryo (HEK 293T)	The cytotoxicity and cell viability of hydrogels, MTT, Live/Dead® assays	-	Mixing	Improvement of physicochemical, morphological and rheological properties	[94]
2-Hydroxypropyltrimethyl ammonium chloride chitosan, polyvinyl alcohol	Nano-hydroxyapatite, sodium citrate dihydrate	Mouse preosteoblast cells MC3T3-E1	Tests of cell viability and proliferation	-	Freezing/thawing technique and immersing process	Improvement of mechanical and tribological properties, biological compatibility	[95]
Polyvinyl alcohol, polyvinyl pyrrolidone	Stick-like TiO2 nanostructures	Human osteosarcoma (HOS; MG-63) cell line	Osteoblast adhesion and proliferation	-	Sol–gel method	Excellent antibacterial efficiency, well cell adhesion and proliferations, bone formation improved	[96]
Glycol, chitosan	Nano-hydroxyapatite	Human sarcoma cell line culture, kidney cell line of a human embryo culture (HEK293T cells) and human bone marrow mesenchymal stem cells (HBMS)	MTT assay, Live/Dead® assays	-	solvent cast and evaporation	Potential bone-related biomedical applications	[97]
Chitosan, β-glycerophosphate disodium salt, gelatin	Bioactive glass nanoparticles	Rat bone marrow mesenchymal stem cells	Cytocompatibility of the hydrogels	Injecting hydrogels into dorsum of Swiss rats	Sol gel method	27% increase in cell viability	[98]
Alginate, polyvinyl alcohol	Chondroitin sulfate loaded zein nanoparticles	Chondrocytes	Degradation studies, chondrocyte culture, Live/Dead® assays, MTS assay, RT-PCR, western blotting	-	Constant stirring and ultrasonication	Chondrocyte improvement, biomimetic matrices upregulating early chondrogenic marker gene (Sox-9) and differentiated genes specific for hyaline cartilage	[99]
Cellulose nanocrystal/dextran hydrogels	Kartogenin and ultrasmall superparamagnetic iron-oxide	Bone marrow-derived mesenchymal stem cells	CCK-8 assay, Live/Dead® assays, gene expression levels	Rabbit articular cartilage	-	Mechanical strength, kartogenin long-term release, support of hyaline cartilage regeneration	[100]