Table 2.
Alliaceae-derived OSCs: effects on in vitro models of osteoclastogenesis and osteoblastogenesis.
| Molecule tested | Experimental in vitro model | Concentration | Main effect | Specific outcomes | Authors | Ref |
|---|---|---|---|---|---|---|
| Hot-water extract and ethanol extracts of Allium hookeri roots | MG-63 cells line | 0.1-0.5-1-5-10-25-50-100 μg/ml | Increased proliferation and osteogenesis | • ↑ viability/proliferation; no cytotoxicity (WST-8 assay) • ↑ ALP activity (pNPP detection) • ↑ collagen (Sirius red assay) • ↑ mineralization (Alizarin Red staining) |
Park et al. | (48) |
| Aqueous and ethanolic extracts of Allium fistulosum | MG-63 cell line | 1-4-8-10-16-32-50-63-125 μg/ml | Increased osteogenesis | • no cytotoxicity (MTT assay) • ↑ ALP activity (ALP assay kit) |
Ryuk et al. | (80) |
| Water solution of onion crude powder | MG-63 cell line | 300 μg/ml | No effect on proliferation or differentiation | • ALP activity similar to control cells (ALP assay kit) • Col I on cell lysate was similar to control cells (4- hydroxyproline quantification) • OCN, OPN in cells surnatants similar to control cells (ELISA) |
Tang et al. | (81) |
| Aqueous and ethanolic extracts of Allium fistulosum | MC3T3-E1 cell line | 1-4-8-10-16-32-50-63-125 μg/ml | Increased proliferation and osteogenesis | Ethanolic extracts: • ↑ viability/proliferation; no cytotoxicity (MTT assay) • ↑ALP activity (ALP assay kit) Water extracts: • no cytotoxicity (MTT assay) • ↑ALP activity (ALP assay kit) |
Ryuk et al. | (80) |
| Water Allium sativum L. extract | Human fetal osteoblast cells | 3D-printed calcium phosphate scaffolds releasing ginger and garlic extract | Increased osteoblast proliferation | • ↑ proliferation (MTT assay) | Bose et al. | (82) |
| Allyl sulfide (AS) * | BMMSCs isolated from Age-associated OP mice’s femurs |
Mice were fed by oral gavage with AS (200 mg/kg) for 3-months | • Rescue of proliferation and osteogenesis • Indirect inhibition of osteoclastogenesis |
• ↑ proliferation as compared to aged mice (MTT assay) • ↑ALP activity (ALP staining), ↑ mineralization (Alizarin red staining), • ↑ RUNX-2 and OCN in cells (western blot) • ↑ OPG and ↓ RANKL in surnatants (ELISA) |
Behera et al. | (83) |
| Allium cepa L. extracts | In vitro bioactivity assay (simulated body fluid) | Chitosan + Allium cepa L. (ChAC) and Chitosan + Allium cepa L. + PLGA (ChPAC) | Improved natural bioactivity of chitosan | • Increased apatite cristals in the surface • Improved Phosphorous/Calcium ratio |
Monárrez-Cordero et al. | (84) |
| Water Allium sativum L. extract | Human osteoclast cells from THP1 monocytes | 3D-printed calcium phosphate scaffolds releasing ginger and garlic extract | Inhibition of osteoclast activity | • ↓ resorption (pit assay) | Bose et al. | (82) |
|
Ethanolic extract of
onion |
RAW 264.7 cell line | 0.1-0.2-0.4 mg/ml | Inhibition of osteoclastogenesis | • no cytotoxicity (MTT assay) • ↓ osteoclasts (TRAP assay) |
Law et al. | (85) |
| Freeze dried onion juice | RAW 264.7 cell line | 0.1-0.2-0.4 mg/ml | Inhibition of osteoclastogenesis | • no cytotoxicity (MTT assay) • ↓ osteoclasts (TRAP assay) |
Law et al. | (85) |
| Water solution of onion crude powder | RAW 264.7 cell line | 15-50-150-300 μg/ml | Inhibition of osteoclastogenesis | • no cytotoxicity (MTT assay) • ↓ osteoclasts (TRAP assay) • ↓ CD51/61 (vitronectin receptor), MMP-9 and TRAP mRNA (RT-PCR) • ↓ ERK, p38 and NF-κB (western blot) |
Tang et al. | (81) |
| Diallyl disulfide (DADS) * | RAW 264.7 cell line | 1-10-100-1000 μg/ml 20-40-60-80-100 μg/ml |
Inhibition of osteoclastogenesis and bone resorption | • ↓ cytotoxicity at concentration higher to 100 μg/ml (CCK-8 assay) • ↓ osteoclast and resorption (TRAP assay PIT assay) • ↓ c-fos, NFATc1, TRAP, MMP9, CTR, CTSK, DC-STAMP, OC-STAMP mRNA • ↓ osteoclast fusion (FAK staining) • ↓ NF-ĸB, p-STAT3, NFATc1, c-FOS (western blot) |
Yang et al. | (86) |
| Alliin * | RAW 264.7 cell line | 0.1-0.5-1-5-10-100 μg/ml | Inhibition of osteoclastogenesis | • No cytotoxicity (CCK-8 assay) • ↓ osteoclasts and resorption (TRAP assay and pit assay) • ↓ c-fos, NFATc1, MMP9, DC-STAMP, OC-STAMP, RANK, TRAP (RT-PCR) • ↓ Nox-1, NFATc1, c-fos (western blot) • ↓ ROS (detection by fluorescent probe) |
Chen et al. | (87) |
| Water solution of onion crude powder | Osteoclast derived from bone marrow cells of femurs of 6-8-week-old Sprague–Dawley rats | 15-50-150-300 μg/ml | Inhibition of osteoclastogenesis | • no cytotoxicity (MTT assay) • ↓ osteoclasts (TRAP assay) |
Tang et al. | (81) |
| Water solution of onion crude powder | Osteoclast derived from long bones of 6-day-old rabbits | 15-50-150-300 μg/ml | Inhibition of bone resorption | • ↓ resorption (pit assay) | Tang et al. | (81) |
| Commercial onion powder (Chia Hui, Taipei, Taiwan) | Osteoclast derived from bone marrow cells of femurs of 6-8-week-old Sprague–Dawley rats | 300 μg/ml | Inhibition of osteoclastogenesis | • ↓ osteoclasts (TRAP assay) • Inhibition of ERK, p38, and NF-κB activation (western blot) |
Tang et al. | (81) |
|
GPCS isolated by bioassay-guided fractionation of
Allium cepa L. Bulbs * |
Osteoclasts derived from femora and tibiae of 2-days-old Wistar Hanlbm rats | 1-10-30 mg/ml 2-4-8 mM |
Inhibition of osteoclast differentiation and activity | • ↓ osteoclast differentiation and resorption by GPCS (TRAP staining and pit assays) | Wetli et al. | (41) |
| Diallyl disulfide (DADS) * | BMMs obtained from the femur and tibia bone marrow of 6-wk-old C57BL/6 mice | 20-40-60-80-100 μg/ml | Inhibition of osteoclastogenesis | • ↓ cytotoxicity at concentration higher to 100 μg/ml (CCK-8 assay) • ↓ osteoclast (TRAP assay) |
Yang et al. | (86) |
| Allyl sulfide (AS)* | BM cells | Cultured under 15% conditioned medium derived from BMMSCs culture of Age-associated OP mouse model (Fed by oral gavage with AS (200 mg/kg) for 3-months) | Inhibition of osteoclastogenesis via a paracrine mechanism | • ↓ osteoclasts (TRAP staining) • ↓ TRAP-5b expression in cells lysates (ELISA) • ↓ NFATc1, CTSK, RANK and OC-STAMP mRNA (RT-PCR) |
Behera et al. | (83) |
Most in vitro studies were conducted by using water or ethanol extracts from Allium edible plants (4 studies, 13 in vitro models; Allium hookeri roots, Allium fistulosum, Allium sativum L., Allium cepa L.); a few used purified OSCs (3 studies, 6 in vitro models; diallyl disulfide (DADS), allyl sulfide (AS), γ-glutamyl-trans-S-1-propenyl-L-cysteine sulfoxide – GPCS, alliin). Most studies showed an increased osteoblast proliferation and osteogenesis and an inhibited osteoclastogenesis. Notably, only the effects of purified OSCs (labeled with * in the table) can be attributable entirely to OSCs. The concentrations tested ranged from 0.1 to 300 μg/ml. Murine in vitro models of osteoclastogenesis: osteoclasts derived from bone marrow of femora and tibiae of rats, rabbits, mice; RAW 264.7 cells. Human in vitro models of osteoclastogenesis: osteoclast cells from human THP1 monocytes. Murine in vitro models of osteoblastogenesis used: MC3T3-E1 (mouse C57BL/6 calvaria cells line); murine bone marrow (BM) cells; bone marrow-derived mesenchymal stem cells (BMMSCs) isolated from age-associated (AG) osteoporosis (OP) mice’s femurs. Murine in vitro models for studying indirect inhibition of osteoclastogenesis: bone marrow-derived mesenchymal stem cells (BMMSCs), bone marrow macrophages (BMM) and murine bone marrow (BM). Human in vitro models of osteoblastogenesis: MG-63 cells line (human osteosarcoma cells line), human fetal osteoblast. Functional assays for osteoclastogenesis used: tartrate-resistant acid phosphatase positive (TRAP staining); pit assay. Functional assays for osteoblastogenesis: alizarin red staining (marker of mineralization), sirius red assay (marker of collagen I), p-nitrophenyl phosphate (pNPP) measurement. Proliferation/viability assays: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell counting kit-8 (CCK-8) cell viability assay, water-soluble tetrazolium-8 (WST-8) assay. Markers of osteoclasts: nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), cathepsin K (CTSK), receptor activator of NF-KB (RANK), osteoclast stimulatory transmembrane protein (OC-STAMP), tartrate-resistant acid phosphatase (TRAP), tartrate-resistant acid phosphatase 5b (TRAP-5b), receptor activator of nuclear factor-κB ligand (RANKL), dendritic cell specific transmembrane protein (DC-STAMP), reactive oxygen species (ROS), calcitonin receptor (CTR), p-signal transducer and activator of transcription 3 (p-STAT3), NADPH Oxidase 1 (Nox-1), c-fos, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), p38, extracellular signal-regulated kinase (ERK), matrix metallopeptidase 9 (MMP-9), CD51/61 (vitronectin receptor). Markers of osteoblastogenesis: collagen I (Col I), osteocalcin (OCN), osteopontin (OPN), runt-related transcription factor 2 (RUNX-2), osteoprotegerin (OPG), alkaline phosphatase (ALP). ↑ means up-regulation; ↓ means down-regulation.