Objective To investigate the variances in the effects of photobiomodulation therapy (PBM) with different wavelength lasers on chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods The second-generation BMSCs were used in the experiment, and laser irradiation treatment was initiated on the first day after passage at a close distance of 7 cm. The experiment was divided into four groups:a control group,a 660 nm group,an 810 nm group, and a 1064 nm group. The control group did not undergo photobiomodulation therapy. The single energy density of laser induction for each group was 4 J/cm2, and the irradiation duration was 15 minutes. After chondrogenic induction culture,the irradiation was repeated every 3 days for 21 days. Cell proliferation activity was evaluated by the CCK-8 method, and glycosaminoglycan (GAG) deposition was detected through toluidine blue staining. The transcription and protein expression levels of chondrogenic marker genes (Aggrecan, Col2A1, GAG) were analyzed by qRT-PCR and Western Blot,respectively. The cell proliferation activity, glycosaminoglycan (GAG) deposition, and the transcription and protein expression levels of chondrogenic marker genes (Aggrecan, Col2A1, GAG)in the three groups under the different wavelengths were compared. Results The CCK-8 results showed that compared with the control group,the proliferation rate of BMSCs under the three wavelengths of laser treatment significantly increased, among which the proliferation-promoting effect of the 1064 nm group was notably superior to that of the 660 nm and 810 nm groups (P＜0.05). The toluidine blue staining revealed that after photobiomodulation therapy,the extracellular matrix metachromatic area of BMSCs was significantly expanded compared to the control group,and the secretion of GAG in each group significantly increased. Compared with the control group, the protein expression levels of Aggrecan and Col2A1 in the photobiomodulation therapy groups were significantly elevated (P＜0.05), and the mRNA expression levels of Aggrecan, Col2A1, and GAG were significantly enhanced (P＜0.05), with the 1064 nm group demonstrating the most significant effect. Conclusions Specific wavelengths (especially 1064 nm) of PBM can significantly optimize the chondrogenic differentiation efficacy of BMSC,providing experimental basis for the optimization of cartilage regeneration strategies in tissue engineering.
Key words
photobiomodulation /
low-level laser /
chondrocyte differentiation /
cell experiment /
mesenchymal stem cell
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