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Table 2 Non-viral gene-delivery based therapeutic approaches for bone and/or cartilage repair

From: Non-viral gene delivery to human mesenchymal stem cells: a practical guide towards cell engineering

(Therapeutic NA)

Non-viral vector

In vitro / in vivo study

Approach

Methods

Ref

pDNA (BMP-2)

PEI polyplexes

In vitro (BMMSCs)/ In vivo (rabbit model)

Bone regeneration

Histological, IHC, and immunofluorescence analyses

[5]

pDNA (BMP-2 and FGF-2)

PEI polyplexes

In vitro (DPMSCs)/ Ex vivo (tooth model)

Bone regeneration

ELISA, PCR, and ICC analyses

[202]

pDNA (BMP-2)

nHA

In vitro BMMSCs)/ In vivo (mice model)

Bone regeneration

ELISA, histological and IHC analyses

[23]

pDNA (BMP-2)

nHA

In vitro (BMMSCs)/ In vivo (rat model)

Bone regeneration

ELISA, histological and IHC analyses

[22]

pDNA (BMP-2, and VEGF)

GET peptide complexes

In vitro (rMSCs) / In vivo (rat model)

Bone regeneration

ELISA, and histological analyses

[178]

pDNA (BMP-2)

TransIT® lipopolyplexes

In vitro (BMMSCs)/ In vivo (rat model)

Bone regeneration

ELISA, histological and IHC analyses

[21]

pDNA (BMP-2)

Poly (amido amine) polyplexes

In vitro (TDMSCs)/ In vivo (mice model)

Bone regeneration

PCR, WB, histological and IHC analyses

[20]

pDNA (BMP-2/6 and BMP-2/7)

Nucleofector®

In vitro (gMSCs)/ In vivo (mice model)

Bone regeneration

ELISA, and histological analysis

[65]

pDNA (BMP-2 and VEGF)

PEI polyplexes and nHA

In vitro (rMSCs) / In vivo (rat model)

Bone regeneration

ELISA, and histological analysis

[69]

pDNA (PDGF B)

PEI polyplexes

In vitro (BMMSCs)/ In vivo (rat model)

Bone regeneration

Histological and IHC analyses

[176]

pDNA (TGF-β1)

Peptide complexes

In vitro (BMMSCs)/ In vivo (rabbit model)

Bone regeneration

ALP assay, PCR, and histological analysis

[18]

pDNA (BMP-2)

PEI polyplexes

In vitro (BMMSCs)

Bone regeneration

PCR, and CC analysis

[177]

pDNA (BMP-2)

nHA (mineral coated)

In vitro (hMSCs)

Bone regeneration

ELISA, and CC analysis

[190]

pDNA (BMP-2/9)

PEI polyplexes

In vitro (BMMSCs) / In vivo (rat model)

Bone regeneration

PCR, histological analysis, and atomic absorption spectroscopy

[218]

PDNA/ (BMP-2 modified)

Peqfect® peptide complexes

In vitro (rAMSCs)

Bone regeneration

ALP assay, ELISA, PCR, and CC analysis

[203]

pDNA (TGF-β1)

Stearate cationic peptide complexes

In vitro (MSCs)

Bone regeneration

ALP assay

[189]

pDNA (OSX-GFP)

bPEI polyplexes

In vitro (hBMMSCs)

Bone regeneration

CC analysis

[174]

pDNA (ephrinB2)

bPEI polyplexes

In vitro (hBMMSCs)

Bone regeneration

PCR, Calcium Liquicolor kit, and blocking peptide assay

[152]

pDNA (BMP-2)

nHA and Lipofectamine 2000®

In vitro (rMSCs)

Bone regeneration

Calcium Liquicolor kit, CC and immunofluorescence analyses

[175]

cDNA (BMP-2)

Lipofectamine 2000® lipoplexes

In vitro (BMMSCs)/ In vivo (mice model)

Bone regeneration

ELISA, and histological analysis

[115]

miRNA (miR-133a)

nHA

In vitro (rMSCs)/ In vivo (rat model)

Bone regeneration

PCR, histological and IHC analyses

[206]

miRNA (miR-122)

Commercial transfection reagents (not specified)

In vitro (rMSCs)/ In vivo (rat model)

Bone regeneration

PCR, WB, histological and IHC analyses

[180]

miRNA (antmiR-138)

Lipofectamine 2000®

In vitro (BMMSCs)/ In vivo (mice model)

Bone regeneration

PCR, WB, histological and IHC analyses

[117]

miRNA (antmiR-138)

Lipofectamine 2000®

In vitro (rat BMMSCs)

Bone regeneration

ALP assay, PCR, and CC analysis

[116]

siRNA (siNoggin)

Lipofectamine 2000® and cationic stereosomes

In vitro (AMSCs)/ In vivo (mouse model)

Bone regeneration

ALP assay, PCR, and histological analysis

[118]

pDNA (RUNX2); siRNA (siOCT3, siOCT4 and siNANOG)

Poly (β-amino ester) polyplexes

In vitro (DPPSCs)

Bone regeneration

ALP assay, PCR, and ICC analysis

[26]

pDNA (BMP-2); siRNA (siNoggin)

C32-122 polyplexes and NA114 lipoplexes

In vitro (BMMSCs)

Bone regeneration

ALP assay, ELISA, PCR, Calcium Reagent kit, and CC analysis

[34]

siRNA (siNoggin); miRNA (miR-20a)

PEI polyplexes

In vitro (hMSCs)

Bone regeneration

CC analysis

[68]

siRNA (siNoggin); miRNA (miR-20a)

PEI polyplexes

In vitro (hMSCs)

Bone regeneration

ALP assay, PCR, Calcium Reagent kit, and CC analysis

[191]

pDNA (BMP-2 and TGF-β3)

nHA and PEI

In vitro (MSCs)

Osteochondral regeneration

ELISA, biochemical, histological and ICC analyses

[19]

pDNA (TGF-β1)

Gelatin microspheres

In vitro (hMSCs)

Chondrogenic differentiation

Biochemical, histological and ICC analyses

[1]

pDNA (TGF-β1)

Pullulan spermine polyplexes

In vitro (rMSCs)

Chondrogenic differentiation

CC analysis

[135]

pDNA (SOX9)

PEI-modified PLGA polyplexes

In vitro (hMSCs)

Chondrogenic differentiation

PCR, biochemical analysis, and immunoblotting

[27]

pDNA (Endostatin)

GP2® lipoplexes

In vitro (cBMMSCs)

Chondrogenic differentiation

ELISA, and CC analysis

[188]

mRNA (SOX9 and MYOD)

3DFectIN® lipoplexes

In vitro (hMSCs)

Chondrogenic and myogenic differentiation

PCR, and ICC analysis

[194]

pDNA (BMP-2 and TFG-β3)

Lipofectamine 2000® lipoplexes

In vitro (hiPSCs)/ In vivo (rat model)

Osteochondral regeneration

PCR, histological and immunochemical analysis

[111]

pDNA (EGFP-C1)

PAA-BA and PEI polyplexes; Lipofectamine 2000® lipoplexes

In vitro (BMMSCs)

Multipotent differentiation

PCR, and ICC analysis

[120]

pDNA (BMP-2 and TGF-β3)

PEI polyplexes, HA nanoparticles, RALA peptide

In vitro (hMSCs)

Osteochondral differentiation

ELISA, PCR, biochemical, CC and ICC analyses

[179]

pDNA (BMP-2 and TGF-β1)

nHA (mineral-coated) and Lipofectamine 2000® lipoplexes

In vitro (hMSCs)

Osteogenic differentiation

Biochemical and CC analyses

[114]

pDNA (BMP-2 and TGF-β3)

CaP nanoparticles or CaP/PEI polyplexes

In vitro (hMSCs)

Osteogenic differentiation

PCR, and CC analysis

[153]

pDNA (BMP-2 and TGF-β1)

Scaffold HA

In vivo (rabbit model)

Osteogenic differentiation

Histological analysis

[186]

miRNA (miR100-5p and miR143-3P)

PEI polyplexes

In vitro (MSCs)

Osteogenic differentiation

PCR, and CC analysis

[67]

miRNA (miR-133a)

nHA

In vitro (hMSCs)

Osteogenic differentiation

PCR, Calcium Liquicolor kit, and ICC analysis

[183]

miRNA (antmiR-138)

Chitosan/tripolyphosphate/HA nanoparticles

In vitro (MSCs)

Osteogenic differentiation

PCR, WB, and CC analysis

[184]

miRNA (miR-218)

bPEI polyplexes

In vitro (hMSCs)

Osteogenic differentiation

ALP assay, PCR, WB, and CC analysis

[182]

  1. Abbreviations. pDNA plasmid DNA, BMP-2, -6, -7, -9 bone morphogenic protein-2, -6, -7, -9, PEI polyethyleneimine, BMMSCs bone marrow derived mesenchymal stem cells, FGF-2 fibroblast growth factor, CC cytochemical, DPMSCs dental pulp mesenchymal stem cells, nHA hydroxyapatite nanoparticles, ELISA enzyme-linked immunosorbent assay, PCR polymerase chain reaction, VEGF vascular endothelial growth factor, rMSCs rat mesenchymal stem cells, GET GAG-binding enhanced transduction, TDMSCs tonsil derived mesenchymal stem cells, gMSCs goat mesenchymal stem cells, TGF-β1 and -β3 transforming growth factor-β1 and -β3, ALP alkaline phosphatase, rAMSCs rat adipose derived mesenchymal stem cells, OSX osterix, GFP green fluorescent protein, bPEI branched polyethyleneimine, cDNA circular DNA, miRNA micro-RNA, siRNA small interfering RNA, RUNX2 core binding factor alpha 2, OCT3 and 4 octamer-binding transcription factor 3 and 4, DPPSCs dental pulp pluripotent stem cells, hMSCs human mesenchymal stem cells, SOX9 sex-determining region Y-type high mobility group box 9, cBMMSCs caprine bone marrow derived mesenchymal stem cells, hiPSCs human induced pluripotent stem cells, MYOD myogenic differentiation protein, EGFP enhanced green fluorescent protein, CaP calcium phosphate, HA hyaluronic acid, PAA-BA poly (amidoamine) with pendant aminobutyl group