Table 3.

Summary of All Investigations for Treatment of Glioblastoma with PAMAM Dendrimer Technology - Inhibition of GB Growth and Proliferation

Investigators	Cytotoxic Cargo	Cell Lines	Results	Cell Viability Measurement	Distribution Analyses
Singh et al (2019)59	Docetaxel	U87MG GL261	Hybrid dendrimer (G4-G3.5) constructs conjugated with docetaxel were increasingly toxic to U87MGMG cells at high concentrations, non-toxic to control cells, and stable in storage over 180 days.	[In vitro] MTT assay & flow cytometry	N/A
Munro et al (2019)61	Curcumin	GL261	D-Cys-Cur and D-Cys transfection of GL261 cells demonstrated similar anti-inflammatory properties. Treatment with both complexes in mice equally prolonged mouse lifespan. No difference in mouse tumor sizes were found between treatment groups.	[In vitro] MTT assay	N/A
Wu et al (2018)77	cMBP	U87MG	MET-targeting cMBP peptides (Den-cMBP10) conjugated to G4 dendrimers showed inhibition of U87MG cell proliferation with reduced pMET, pAKT, and pERK1/2 expression levels. Glioma volume size decreased significantly with Den-cMBP10 and median mouse survival increased by 59%.	[In vitro] TUNEL assay	[In vivo] MRI
Uram et al (2018, 2019)78,79	Celecoxib Fmoc-L-Leucine	U-118 SCC-15 HaCaT	Conjugated dendrimer delivery of both celecoxib and Fmoc-L-Leucine at a ratio of 1:1 showed significant cytotoxicity in U-118 cells. Decreased cell viability, mobility, and proliferation in U-118 cells with administration of a lower dose of G3-BCL (1–2 μM)	[In vitro] NR assay [In vitro] Apo-ONE®Homogenous Caspase-3/7 Assay	[In vitro] Fluorescence confocal microscopy
Choi et al (2017)66	Quercetin Acetazolamide	U251	Quercetin conjugated PEG-PLGA micelles induced minimal cell death and reduced nitric oxide release. Acetazolamide conjugated PEG-PLGA micelles showed increased cell death.	[In vitro] Hoechst stain & spheroid cultures	[In vitro] UV-absorbance spectrometry
Bae et al (2017)71	Apoptin	U87MG	PAMAM-H-R and PAMAM-H-K showed increased transfection efficiency compared to PAMAM. Decreased cell cytotoxicity with PAMAM-H-R/pJDK-apoptin use and greater apoptosis induction and loss of mitochondrial membrane potential than PAMAM-H-K/pJDK-apoptin.	[In vitro] EZ-Cytox cell viability assay, Caspase 3 activity assay, & cell cycle distribution	[In vitro] flow cytometry & fluorescence confocal microscopy
Pedro-Hernandez et al (2017)72	Ibuprofen	U251 PC-3 K-562 HCT-15 MCF-7 SKLU-7 MDA-MB-231	Ibuprofen conjugated to the resorcinarene-PAMAM dendrimers resulted in high inhibition of cell growth and cytotoxicity in all cell lines. Dendrimer conjugation and modification potency: ethylphenyl group with 16 ibuprofen moieties > dodecyl group with 16 ibuprofen moieties > dodecyl group with 8 ibuprofen moieties > ethylphenyl group with 8 ibuprofen moieties.	[In vitro] Protein-binding dye sulforhodamine B cytotoxic assay	[In vitro] hydrolysis release analysis & fluorescence microscopy
Janiszewska et al (2016)73	siRNA	U87MG C6	PLL dendrimers protect siRNA from degradation and inhibited cell proliferation of GB cell lines; PLL dendrimers mediate cell toxicity with ROS production after 24 hrs with mitochondrial depolarization to inhibit GB cell proliferation.	N/A	[In vitro] fluorescence microscopy
Bae et al (2016)70	Apoptin	GBL-14 GBL-37	PAMAM-H-R shows better transfection efficiency and higher expression of apoptin vs PAMAM and PAMAM-H-K dendrimers. 36 hr transfection in tumor cells with PAMAM-H-R/pJDK-apoptin resulted in cell death and no apoptosis in dermal fibroblasts with loss of mitochondrial membrane potential and depletion of cell glutathione levels.	[In vitro] EZ-Cytox cell viability assay kit, LDH assay, & glutathione assay	[In vitro] confocal microscopy & FACS
Lesniak et al (2016)90	Salicylic acid	U87MG	G5 PAMAM dendrimers conjugated to salicylic acid delivered to U87MG GB-bearing mice. CEST and MRI imaging showed 50% of the tumor images contained the G5 dendrimer construct.	N/A	[In vivo] chemical exchange saturation transfer (CEST) MRI
Bai et al (2013)89	IFN-β	U87MG	G4 PAMAM-R/pORF dendrimers packaged with IFN-β inhibited cancer cell growth in vitro by 27%. Xenografts of cancer cells with dendrimer construct inhibited cancer cell growth.	[In vitro] MTT assay [In vivo] TUNEL assay	[In vitro] ELISA (GFP and luciferase)
Perez et al (2011)34	siRNA	T98G J774	G7 dendrimers packaged with siRNA administered with an endocytotic inhibitor. Dendriplexes were taken up by clathrin-dependent endocytosis and caveolin-mediated endocytosis in J774 cells and by cholesterol, caveolin, and actin cytoskeleton pathways in T98G cells.	N/A	[In vitro] Trypan blue and FITC-labelled fluorescence microscopy
Ren et al (2010)55	Taxol miR-21 inhibitor	U251	PAMAM dendrimers packaged with taxol and miR-21 inhibitors enhanced cell apoptosis in U251 and LN229 (control) cells.	[In vitro] MTT assay [In vivo] annexin V/PI staining and flow cytometry	[In vitro] microscopy of transwell chambers
McNerny et al (2009)57	Methotrexate	U87MG HUVEC	PAMAM dendrimers conjugated with c(RGDyK) packaged with methotrexate inhibited tumor growth in vitro.	[In vitro] XTT assay	[In vitro] flow cytometry [In vivo] confocal microscopy
Kaneshiro et al (2009)58	Doxorubicin Luciferase siRNA	U87MG	G3 Poly(L-lysine) dendrimers with a silsesquioxane cubic core and c(RGDfK) conjugate were packaged with doxorubicin and siRNA. Successfully inhibited U87MG-Luc cell growth from doxorubicin-induced cytotoxicity and siRNA effects.	[In vitro] MTT assay	[In vitro] Cy3-siRNA confocal microscopy