Table 3.
Study Design and Biochemical, Histological, and Behavioral Outcomes for In Vivo Studies.
| First Author, Year | Operating
Mode Treatment Duration (No. of Sessions) |
Experimental Design | OA Model | Animal
Type Gender Animal Race Age (months) Weight (g) |
Biochemical Outcomes | Histological Outcomes | Behavioral Outcomes | |||
|---|---|---|---|---|---|---|---|---|---|---|
| ECM Synthesis/Degradation | Inflammatory and Pain Markers | Osteoarthritis Grade | Morphometric Analysis | Cartilage Organization | ||||||
| Trevisan, 2020 21 | Continuous 3 times per week (12) |
G1: OA (n = 10) G2: OA + LT (n = 10) |
ACL transection | 20 Rats Male Wistar 2 months 150 g |
↑ COL II [G2 vs G1]
(immunoexpression) ↑ TGF-β (growth factor) [G2 vs. G1] (immunoexpression) |
NA | ↓ OARSI score [G2 vs. G1] |
NA | Abnormal chondrocyte orientation and proliferation. Few irregularities and fibrillation after LT. | No effect on gait performance |
| Yamada, 2020 9 | Pulsed 3 times per week (8) |
G1: Saline injection (n =
10) G2: Saline + LT at 18 J/cm2 (n = 10) G3: OA (n = 10) G4: OA + LT at 6 J/cm2 (n = 10) G5: OA + LT at 18 J/cm2 (n = 10) |
Intra-articular injection of MIA | 50 Rats Male Wistar 3 months 250 g |
NA | ↓ Lipid oxidation [G4/G5 vs. G2] in spinal
cord and [G4 vs. G2] in blood
serum ↓ Protein carbonyl [G5 vs. G2] in spinal cord ↑ SOD activity [G4/G5 vs. G2] in brainstem ↓ Nonprotein thiol [G4/G5 vs. G2] in blood serum ↑ Nonprotein thiol [G4/G5 vs. G2] in brainstem ↓ MPO activity [G5 vs. G2] in intra-articular lavage ↑ MPO activity [G4/G5 vs. G2] in blood Serum ↓ IL-1β, TNF-α, IL-6 [G5 vs. G2] No differences in IL-10 (protein expression by ELISA) |
NA | NA | NA | ↑ Mechanical Hyperplasia ↑ Spontaneous pain [G5 vs. G2] |
| Li, 2020 8 | NR daily (7) |
G1: Saline injection (n = 8) G2: OA (n = 8) G3: OA + LT (n = 8) G4: OA + sham LT (n = 8) |
Intra-articular injection of MIA | 32
Rats Male Sprague-Dawley NR 220-250 g |
↓ MMP-3 [G3 vs. G2/G4] (immunoexpression) |
↓ TNF-α, ↓ IL-1β, ↓ IL-6 [G3 vs. G2/G4] (protein expression by ELISA) |
↓ OARSI score [G3 vs. G2/G4] |
NA | LT inhibited cartilage destruction and proteoglycan loss. | ↑ Mechanical hyperalgesia ↑ Weightbearing [G3 vs. G2/G4] |
| Balbinot, 2019 7 | Continuous Daily (15) |
G1: OA (n = 9) G2: OA + LT (n = 10) |
Intra-articular injection of MIA | 19 Rats Male Wistar 3 months 355 ± 22 g |
↑ PGs ↑ Chondrocyte content [G2 vs. G1] (toluidine blue staining) |
↓ Reactive astrogliosis in spinal
cord (immunoexpression) [G2 vs. G1] |
NA | ↑ Chondrocyte content [G2 vs. G1] (toluidine blue staining) |
NA | No effect on gait performance; ↑ Weightbearing; ↑ Mechanical Hyperplasia [G2 vs. G1] |
| Stancker, 2018 22 | Continuous Daily (7) |
G1: Culture medium injection (n =
10) G2: OA (n = 10) G3: OA + LT (n = 10) G4: OA + ADSCs (n = 10) G5: OA + ADSCs + LT (n = 10) |
Intra-articular injection of papain solution | 50 Rats Male Wistar 3 months 250-300 g |
↑ COL II [G3/G4 vs. G2] [G5 vs. G2/G4] ↓ MMP-2 and ↓ MMP-13 [G3/G4/G5 vs. G2] No differences among treated groups. (Protein expression by Western blot) ↓ MMP-1, ↓ MMP-2, ↑ TIMP-1 and ↑ TIMP-2 [G3/G4 vs. G2] [G5 vs G2/G3/G4] (Gene expression by PCR) |
↓ IL-1-β and ↓ TNF-α [G3/G4 vs. G2] [G5 vs. G2/G4] ↓ IL-6 [G3/G4 vs. G2] [G5 vs. G2/G3/G4] ↑ IL-10 [G3 vs. G2] [G5 vs. G2/G4] (Gene expression by PCR) |
NA | NA | NA | NA |
| Sanches, 2018 23 | Continuous 3 times a week (29) |
G1: OA (n = 10) G2: OA + LT (n = 10) G3: OA + CS/GL (n = 10) G4: OA + LT + CS/GL (n = 10) |
ACL transection | 40 Rats Male Wistar 2 months 150 g |
↑ COL II [G4 vs. G1]
No differences among treated groups. (immunoexpression) |
↓ IL-1β and ↑ IL-10 [G4 vs. G1]
No differences among treated groups. (immunoexpression) |
↓ OARSI score [G3/G4 vs. G1] No differences among treated groups |
↓ Density of chondrocytes [G2/G3/G4 vs. G1] No differences among treated groups. No difference in cartilage thickness |
Degradation, fibrillation, hypercellularity and chondrocytes disorganization among treated groups. | NA |
| Assis, 2018 24 | Continuous NR |
G1: OA (n = 10) G2: OA + aerobic exercise (n = 10) G3: OA + aquatic exercise (n = 10) G4: OA + aerobic exercise + LT (n = 10) G5: OA + aquatic exercise + LT (n = 10) |
ACL transection | 50 Rats Male Wistar 1.5 months 150 ± 11.2 g |
No effect on COL I ↑ COL II [G2/G3/G4/G5 vs. G1] No differences among treated groups. (immunoexpression) |
↑ IL-10 [G2/G3/G4/G5 vs. G1]
↑ TGF-β [G2/G4 vs. G1] No differences among treated groups. (immunoexpression) |
↓ OARSI score [G2/G3/G4/G5 vs. G1] No differences among treated groups. |
NA | Abnormal chondrocyte orientation and proliferation, few irregularities and fibrillation among treated groups. | NA |
| Tomazoni, 2017 25 | Continuous 3 times a week (24) |
G1: No intervention (n = 6) G2: OA (n = 6) G3: OA + exercise (n = 6) G4: OA + NSAID (n = 6) G5: OA + LT (n = 6) G6: OA + exercise + NSAID (n = 6) G7: OA + exercise + LT (n = 6) G8: OA + NSAID + LT (n = 6) G9: OA + exercise + LT + NSAID (n = 6) |
Intra-articular injection of papain solution | 54
Rats Male Wistar NR 200-250 g |
NA | ↓ IL-1β [G5 vs. G2/G3/G4/G6G9] [G7 vs. G6/G9] No effect on IL-6 and TNF-α (gene expression by PCR) ↓ IL-1β [G4/G5/G7/G8 vs. G2] [G5 vs. G2/G7] [G7 vs. G6/G9] ↓ IL-6 [G3/G4/G5/G6/G7/G8/G9 vs. G2] [G5/G7 vs. G4/G6] ↓ TNF-α [G3/G4/G5/G6/G7/G8/G9 vs. G2] [G5/G7 vs. G3/G6] (Protein expression by ELISA) |
NA | NA | NA | NA |
| Mahmoud, 2017 26 | Pulsed 2-3 times a week (10) |
G1: OA (n = 6) G2: OA + US (n = 6) G3: OA + LT (n = 6) |
Injection of saline solution of Lev | 18 Rats Male Albino 3 months 150-200 g |
NA | ↓ IL-1β [G2/G3 vs. G1]
↓ IL-6 [G2 vs. G1] ↑ IL-10 and ↓ INF-γ [G2 vs. G1/G3] No effect on TNF-α (Protein expression by ELISA) |
NA | ↓ Mankin scores [G2/G3 vs. G1] |
Similar histological evidences among treated groups: smooth surface without irregularities. | ↑ Knee maximum extension [G2/G3 vs. G1] |
| Tomazoni, 2016 28 | Continuous 3 times a week (24) |
G1: No intervention (n = 6) G2: OA (n = 6) G3: OA + LT (n = 6) G4: OA + exercise (n = 6) G5: OA + Diclo (n = 6) G6: OA + exercise + LT (n = 6) G7: OA + Diclo + LT (n = 6) G8: OA + exercise + Diclo (n = 6) G9: OA + exercise + LT + Diclo (n = 6) |
Intra-articular injection of papain solution | 54
Rats Male Wistar NR 200-250 g |
↓ MMP-3 [G3/G5/G7/G9 vs. G2] No differences among treated groups. ↓ MMP-13 [G3 vs. G2/G7/G6] [G4/G5/G8/G9 vs. G2/G7] (gene expression by PCR) |
↓ Inflammatory cells number [G3/G5 vs. G2/G4/G6/G7/G8/G9] (intra-articular wash counting) ↑ MPO activity [G3/G6 vs. G2/G4/G5/G7] (synovial supernatant quantification) |
NA | NA | LT and exercise alone promoted a homogeneous chondrocytes distribution. Remaining treated groups presented chondrocytes distributed heterogeneously. | NA |
| de Oliveira, 2016 27 | Continuous NR |
G1: No intervention (n = 18) G2: OA (n = 18) G3: OA + LT (n = 18) |
Intra-articular injection of papain solution | 54 Rats Male Wistar 3 months 250-300 g |
NA | ↓ TNF-α [G3 vs. G2]
(Protein expression by ELISA) |
NA | NA | NA | ↑ Mechanical hyperalgia [G3 vs. G2] |
| Assis, 2016 29 | Continuous 3 times a week (24) |
G1: No intervention (n = 10) G2: OA (n = 10) G3: OA + aerobic exercise (n = 10) G4: OA + LT (n = 10) G5: OA + aerobic exercise + LT (n = 10) |
ACL transection | 50 Rats Male Wistar 1.5 months 150 g |
↓ MMP-13 [G3/G4/G5 vs. G2]
No differences among treated groups. (immunoexpression) |
↓ IL-1β [G3/G4/G5 vs. G2]
No differences among treated groups. ↓ Caspase-3 [G3/G4 vs. G2] [G5 vs. G2/G3/G4] (immunoexpression) |
↓ OARSI score [G3/G4/G5 vs. G2] No differences among treated groups. |
No differences in chondrocytes density ↑ Cartilage thickness [G3/G4/G5 vs. G2] No differences among treated groups. |
Few signs of fibrillation and irregularities, moderate number of chondrocytes and organization among treated groups. | NA |
| Milares, 2016 30 | Continuous 3 times a week (24) |
G1: OA (n = 10) G2: OA + exercise (n = 10) G3: OA + LT (n = 10) G4: OA + exercise + LT (n = 10) |
ACL transection | 40 Rats Male Wistar 1.5 months 150 g |
↓ MMP-13 [G4 vs. G1]
No differences among treated groups. (immunoexpression) |
↓ IL-1β and ↓ Caspase-3 [G2/G4 vs. G1] No differences among treated groups. (immunoexpression) |
↓ OARSI score [G2/G3/G4 vs. G1] No differences among treated groups. |
↓ Chondrocytes density and ↑ Cartilage
thickness [G2/G3/G4 vs. G1] No differences among treated groups. |
Less tissue degradation, marks of fibrillation and irregularities, and chondrocytes organization among treated groups. | NA |
| Mangueira, 2015 31 | Continuous NR |
G1: Saline injection (n = 9) G2: OA (n = 9) G3: OA + LT at 660 nm (n = 10) G4: OA + LT at 780 nm (n = 10) |
Intra-articular injection of collagenase | 36
Rats Male Wistar NR 220-260 g |
↑ COL III area [G3 vs. G2/G4] ↑ COL II area [G3/G4 vs. G2] (Picrosirius red staining) |
NA | NA | NA | NA | NA |
| Dos Santos, 2014 32 | Continuous NR |
G1: No intervention (n = 5) G2: OA (n = 5) G3: OA + PMT at 2 J (n = 5) G4: OA + LT at 4 J (n = 5) |
Intra-articular injection of papain solution | 20 Rats Male Wistar 3 months 250-300 g |
NA | ↓ Neutrophils and ↓ Macrophages [G3/G4 vs. G2] [G3 vs. G4] (Intra-articular wash counting) ↓ IL-1β and ↑ IL-10 [G3 vs. G2] No differences among treated groups. ↓ TNF-α [G4 vs. G2/G3] [G3 vs. G2] ↓ IL-6 [G4 vs. G3] (Gene expression by PCR) |
NA | NA | Similar histological evidences among treated groups: low intensity acute inflammation and normal articular surface. | NA |
| Wang, 2014 34 | Continuous 3 times a week (6, 12, 18, or 24) |
G1: OA (n = 80) G2: OA + LT (n = 80) |
ACL transection | 160 Rabbits NR New Zealand 6 months 3500± 800 g |
↑ COL II, ↑ ACAN, ↑ TGF-β, ↑ MMP-1 and ↑ MMP-13 after 8
weeks [G2 vs. G1]
↑ TIMP-1 and ↑ MMP-3 after 6 and 8 weeks [G2 vs. G1] No effect on IGF-1, BMP-2 and BMP-7 (gene expression by PCR) |
↓ IL-1β after 6 and 8 weeks [G2 vs. G1] (gene expression by PCR) |
↓ OA score after 6 and 8 weeks [G2 vs. G1] |
NA | LT improved cartilage damage and erosion at all locations. | ↑ Weight bearing after 6 and 8 weeks [G2 vs. G1] |
| Dos Santos, 2014 33 | Continuous 5 times a week (15 or 30) |
G1: Control (n = 20) G2: OA (n = 20) G3: LT at 10 J/cm2 (n = 20) G4: LT 50 J/cm2 (n = 20) |
ACL transection | 80 Rats Male Wistar 3 months 300 ± 20 g |
No differences in COL II organization and
intensity ↓ MMP-13 after 8 weeks [G3/G4 vs G2] No differences among treated groups (immunoexpression) |
No effect on TNF-α and
IL-1β (immunoexpression) |
NA | ↓ Cell number after 8 weeks [G3/G4 vs. G2] No differences among treated groups No effect of cartilage thickness |
G4 showed chondrocytes disorganization and intense presence of cells, while G3 presented slight fibrillation and moderate presence of cells after 8 weeks of LT. | NA |
| Bublitz, 2014 35 | Continuous 5 times per week (15) |
G1: OA (n = 10) G2: LT at 10 J/cm2 (n = 10) G3: LT at 50 J/cm2 (n = 10) |
ACL transection | 30 Rats Male Wistar 3 months 300 ± 20 g |
No effect on MMP-13
expression (immunoexpression) |
No effect on IL-1β
expression (immunoexpression) |
NA | No effect on chondrocytes number. ↓ Cartilage area [G2/G3 vs. G1] ↓ PGs reduction score [G2/G3 vs. G1] (safranin O staining) No differences among treated groups |
G2 presented more tissue and chondrocytes organization with no fibrillation in comparison to G1 and G2. G3 presented tissue disorganization in comparison to G2, but better organized than G1. | NA |
| Alves, 2014 36 | Continuous 3 times a week (4, 7, or 10) |
G1: No intervention (n = 15) G2: OA (n = 15) G3: LT at 50 mW (n = 15) G4: LT at 100 mW (n = 15) |
Intra-articular of papain solution | 60 Rats Male Wistar 3 months 250-300 g |
↓ COL I and ↑ COL II [G3/G4 vs G2] (Picrosirius red staining) ↓ MMP-2 and ↓ MMP-9 after 7 and 14 days [G3/G4 vs G2] ↓ MMP-2 and ↓ MMP-9 after 21 days [G3 vs G2] (Protein expression by Western blot) No differences among treated groups |
NA | NA | NA | After 21 days, G3 showed tissue repair, but fewer fibroblast. G4 presented a thick synovial membrane and tissue repair. | NA |
| Oliveira, 2013 37 | Continuous 5 times a week (15 or 30) |
G1: No intervention (n = 20) G2: OA (n = 20) G3: LT at 10 J/cm2 (n = 20) G4: LT at 50 J/cm2 (n = 20) |
ACL transection | 80 Rats Male Wistar 3 months 300 ± 20 g |
No effect on MMP-13 ↑ COL I after 5 weeks [G3 vs. G2] ↓ COL I after 8 weeks [G3/G4 vs. G2] (immunoexpression) No differences among treated groups |
No differences in TNF-α and
IL-1β (immunoexpression) |
No effect on Mankin score | ↓ Chondrocytes number after 8 weeks [G3/G4 vs. G2] No differences among treated groups No effect on cartilage thickness. |
After 8 weeks, G3 showed a better tissue organization compared with G2, moderate number of cells, slight fibrillation and irregularities. G4 exhibited more disorganized tissue compared to G3, moderate presence of chondrocytes and fibrillation. | NA |
| Alves, 2013 38 | Continuous Once |
G1: No intervention (n = 15) G2: OA (n = 15) G3: LT at 50 mW (n = 15) G4: LT at 100 mW (n = 15) |
Intra-articular injection of papain solution | 60 Rats Male Wistar 3 months 250-300 g |
NA | ↓ IL-1β and ↓ IL-6 [G3 vs. G2]
↓ TNF-α [G4 vs. G2/G3] (gene expression by PCR) ↓ Neutrophils [G3/G4 vs. G2] ↓ Macrophages [G3 vs. G2/G4] (Intra-articular wash counting) |
NA | NA | G3 presented tissue with fibroblast and discrete inflammatory cells, while G4 showed acute inflammatory infiltrate, red blood cells and hyaline material presence. | NA |
| Kim, 2013 39 | Pulsed 5 times per week (15) |
G1: Saline injection (n =
10) G2: OA (n = 10) G3: LT (n = 10) |
Intra-articular injection of MIA | 30
Rats Male Sprague-Dawley NR 150-160 g |
NA | ↓ TNF-α, ↓ IL-1β and ↓ IL-6 [G3 vs. G2] (protein expression in blood by ELISA) |
NA | NA | NA | NA |
| Kim, 2013 40 | Pulsed 5 times per week (15) |
G1: Saline injection (n =
10) G2: OA (n = 10) G3: LT (n = 10) |
Intra-articular injection of MIA | 30
Rats Male Sprague-Dawley NR 150-160 g |
NA | NA | NA | NA | NA | ↑ Mechanical hyperplasia after 14 and 21 days ↑ Weightbearing after 21 days [G3 vs. G2] |
| Lin, 2012 41 | Continuous 5 times a week (10) |
G1: No intervention (n = 8) G2: OA (n = 8) G3: LT (n = 8) |
ACL transection | 24 Rabbits male/female (50%-50%) New Zealand 3 months 2,000-2,500 g |
NA | ↓ Caspase-3 [G3 vs. G2]
No effect on Caspase-8 (immunoexpression) |
↓ Markin score [G3 vs. G2] |
NA | After LT, the tissue presented a smoother surface with slight disorganization. | NA |
| Da Rosa, 2012 42 | Continuous Daily (7, 14, or 21) |
G1: OA (n = 12) G2: LT at 660 nm (n = 12) G3: LT at 808 nm (n = 12) |
Intra-articular injection of papain solution | 36 Rats Male Wistar 3 months 250-300 g |
NA | No effect on the number of inflammatory
cells. (immunoexpression) |
NA | NA | ↑ Newly formed vessels after 7 days [G2 vs. G1] ↓ Fibrosis intensity after 14 days [G2/G3 vs. G1] ↓ Fibrosis intensity after 21 days [G3 vs. G1] G3 presented better results regarding the formation of epithelium and new blood vessels. |
NA |
| Oshima, 2011 43 | Pulsed 5 times a week (25) |
G1: OA (n = 7) G2: LT (n = 7) |
ACL transection | 14 Rabbits Female New Zealand 9-15 months 3,500-4,500 g |
↑ COL II [G2 vs. G1]
(gene expression by PCR) No effect on ACAN, MMP-3, and MMP-13 |
↓ TNF-α [G2 vs. G1]
(gene expression by PCR) No effect on IL-1β |
↓ OA grade [G2 vs. G1] (Gross appearance analysis) |
NA | NA | NA |
| Zhao, 2011 44 | NR Every other day (15) |
G1: OA (n = 10) G2: OA + sham LT (n = 10) G3: Laser 10.6 µm + 650 nm (n = 10) G4: LT at 650 nm (n = 10) G5: Laser 10.6 µm (n = 10) |
Naturally after extreme exercise | 50 Mice male/female (50%-50%) C57 Black 5 months 20-25 g |
NA | NA | ↓ Mankin score [G3 vs. G2] |
NA | NR | NA |
| Kamali, 2007 45 | Pulsed Twice per week (8, 16, or 32) |
G1: Control (n = 6-8) G2: LT (n = 6-8) |
Osteo-chondral defect (5 × 4 mm) | 41 Rabbits NR Dutch White 4 months 2000 ± 300 g |
NA | NA | NA | No differences in cartilage thickness. ↑ Cartilage stiffness (biomechanical analysis) after 8 weeks [G2 vs. G1] |
NA | NA |
| Lin, 2006 46 | Continuous 3 times a week (24) |
G1: Early-stage OA; (n =
3-4) G2: Intermediate-stage OA (n = 3-4) G3: Late-stage OA (n = 3-4) G1s, 2s, 3s: LT (n = 3-4) G1c, 2c, 3c: Control (n = 3-4) |
Intra-articular injection of papain solution | 78
Rats Female Wistar NR 320-350 g |
NA | NA | ↓ OA grade [G1s vs. G1c] [G2s vs. G2c] |
NA | G1s and G1c showed fibrillation. G2s and G2c presented chondrocyte enlargement. G3s and G3c exhibited deep fibrillation and partial to total cartilage loss. | NA |
| Cho, 2004 47 | Pulsed Daily (15 or 30) |
G1: OA (n = 5) G2: LT for 2 weeks (n = 5) G3: LT for 4 weeks (n = 5) G4: OA for 2 weeks without LT (n = 5) G5: OA for 4 weeks without LT (n = 5) |
Intra-articular injection of H2O2 | 28 Rabbits NR New Zealand 10 months 2,500-3,000 g |
NA | NA | NA | NA | After 4 weeks of LT, chondrocytes disorganization was observed. | NA |
NR, not reported; NA, not applicable; G, group; LT, light therapy; OA, osteoarthritis; ACL, anterior cruciate ligament; MIA, monosodium iodoacetate; Lev, levofloxacin; COL I, II, and III, collagen type I, II, and III; MMP, matrix metalloproteinase; TIMP, tissue inhibitors of metalloproteinases; IL, interleukin; TNF-α, tumor necrosis factor-α; INF-γ, interferon gamma; ACAN, aggrecan; PGs, proteoglycans; OARSI, Osteoarthritis Research Society International; PCR, polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay; ADSCs, adipose-derived stem cells; US, ultrasounds; NSAID, nonsteroidal anti-inflammatory drug; Diclo, sodium diclofenac; SOD, superoxide dismutase; MOP, myeloperoxidase.