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Table 1 Studies using KPs for osteoporosis prevention and treatment

From: Potential of Kefir-Derived Peptides, Probiotics, and Exopolysaccharides for Osteoporosis Management

Disease type

Experimental models & Intervention regimens (oral route)

Intervention outcomes

Postmenopausal osteoporosis [8]

In vivo (OVX SD rats): 164, 328, and 656 mg KPs/kg BW/day for 12 weeks

In vitro (Caco-2 cells): 400 μg/ml

1. Serum:↓CTX

2. H&E (femur): ↓pathological score

3. μ-CT (femur): ↑BMD, ↑BV/TV, ↑Tb.N, ↑Tb.Th, ↓Tb.Sp

4. Nanoindentation (femoral cortical bone): ↑hardness, ↑elastic modulus

5. Caco-2 cells: ↑TRPV6-dependent Ca2+ influx

Postmenopausal osteoporosis [15]

In vivo (OVX C57BL/6 J mice): 100 mg KPs/kg BW/day, or the combination of KPs and 10 mg CaCO3/kg BW/day for 8 weeks

1. ↓Kidney-surrounding fat

2. μ-CT (femur): ↑BMD, ↑BV/TV, ↑Tb.N, ↓Tb.Sp

3. Nanoindentation (femoral cortical bone): ↑hardness, ↑elastic modulus, ↓residual indentation area

4. 16S rRNA analysis: KPs mildly modulate the structure of gut microbiota in OVX mice

Osteoporosis

[14]

Double-blinded human trial (n = 40):

Control: CaCO3, 1500 mg/day

Treatment: CaCO3 + KPs, 1600 mg/day, for 6 months

1. Serum:↓β-CTX, ↑OC, ↑PTH

2. CT (hip): ↑BMD

Vitamin C deficiency-induced osteoporosis [16]

In vivo (♂AKR1A1-KO mice): oral administration of 164 and 656 mg KPs/kg BW/day for 12 weeks

In vitro (BMMs, BMMSCs): 0 ~ 20 μg/ml KPs

1. Serum:↓ROS, ↓MDA, ↓IL-1β, ↓IL-6, ↓TNF-α, ↓CTX-1, ↓RANKL, ↑SOD, ↑catalase, ↑P1NP, ↑OC, ↑OPG

2. μ-CT (femur): ↑BMD, ↑BV/TV, ↑Tb.N, ↓Tb.Sp

3. Nanoindentation (femoral cortical bone): ↑hardness, ↑elastic modulus

4. OC differentiation: ↑ALP, ↑bone mineralization, ↑Collagen I, ↑β-catenin, ↑RUNX2, ↑BMP-2, ↑NFATc1, ↑HO-1, ↓p53

5. Osteoclast differentiation: ↓TRAP, ↓bone resorption, ↓c-Src, ↓cathepsin K, ↓NFATc1, ↓c-Fos

6. Intracellular signaling: ↓p38, ↓NF-κB, ↓Akt, ↓PLCγ2, ↓CREB-1

Hemophilia-induced osteoporosis [17]

In vivo (♂ Factor VIII-KO mice): oral administration of 164, 328, and 656 mg KPs/kg BW/day for 8 weeks

In vitro (BMMs): 10, 20, and 40 μg/ml KPs

1. Serum: ↓CTX-1, ↓RANKL, ↓IL-6, ↑OPG

2. H&E (femur): ↓pathological lesion, ↓TRAP

3. μ-CT (femur): ↑BMD, ↑BV/TV, ↑Tb.N, ↓Tb.Sp

4. μ-CT (lumbar): ↑Tb.N, ↓Tb.Sp

5. Nanoindentation (femoral cortical bone): ↑hardness, ↑elastic modulus

6. Osteoclast differentiation: ↓TRAP + osteoclasts

Atherosclerosis-induced vascular calcification and osteoporosis [18]

In vivo (♂ ApoE-KO mice with a high-cholesterol diet): oral administration of 328 and 656 mg KPs/kg BW/day for 13 weeks

1. Serum: ↓total cholesterol, ↓ox-LDL, ↓MDA, ↓TNF-α, ↓CTX-1, P1NP ↑

2. Aortic arc (staining): ↓lipid, ↓collagen, ↓inflammation

3. Aortic arc (Western blot): ↓TNF-α, ↓IL-1β

4. μ-CT (femur): ↑BMD, ↑BV/TV, ↑Tb.N, ↑Tb.Th, ↓Tb.Sp, ↓SMI

Femoral fracture [26]

In vivo (♂SD rats with femur-fracture surgery

): oral administration of 400 mg KPs/kg BW/day for 4 weeks

In vitro (BMMSCs): 0 ~ 80 μg/ml KPs

In vitro (MC3T3-E1): 400 μg/ml KPs

1. Serum: ↑OC, ↑P1NP

2. X-ray (fracture site): fracture line completely vanished, full remodeling of cortex, ↑Lane & Sandhu score

3. Staining (fracture site): ↓cartilaginous callus, ↑calcified fibrocartilage

4. mRNA (fracture site): ↑Col1a1, ↑ALP, ↑SPP1, ↑VEGFA, ↑Cox-2

5. MC3T3-E1 cultures (mRNA): ↑Col1al, ↑ALP, ↑M-CSF, ↑Phospho1

6. BMMSC cultures: ↑bone mineralization

  1. Abbreviations: bone marrow macrophage (BMM), bone marrow mesenchymal stem cell (BMMSC), C-terminal telopeptides of type I collagen (CTX), bone mineral density (BMD), bone volume (BV/TV), trabecular number (Tb.N), trabecular separation (Tb.Sp), trabecular thickness (Tb.Th), skeletal muscle index (SMI), parathyroid hormone (PTH), reactive oxygen species (ROS), malondialdehyde (MDA), oxidized low density lipoprotein (ox-LDL), superoxide dismutase (SOD), receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), osteocalcin (OC), procollagen type 1 amino-terminal propeptide (P1NP), alkaline phosphatase (ALP), interleukin (IL), tumor necrosis factor (TNF), nuclear factor-kappa B (NF-κB), Runt-related transcription factor 2 (RUNX2), bone morphogenetic protein-2 (BMP-2), nuclear factor of activated T cells 1 (NFATc1), heme oxygenase 1 (HO-1), Tartrate-resistant acid phosphatase (TRAP), phospholipase C gamma 2 (PLCγ2), cAMP response element-binding protein 1 (CREB-1), alpha-1 type I collagen (Col1a1), secreted phosphoprotein 1 (SPP1), vascular endothelial growth factor A (VEGFA), cyclooxygenase 2 (Cox-2), macrophage colony-stimulating factor (M-CSF), phosphoethanolamine/phosphocholine phosphatase 1 (Phospho1)