Laser-activated autologous adipose tissue-derived stromal vascular fraction restores spinal cord architecture and function in multiple sclerosis cat model

Abstract

Background Multiple sclerosis (MS) is the most frequent non-traumatic neurological debilitating disease among young adults with no cure. Over recent decades, efforts to treat neurodegenerative diseases have shifted to regenerative cell therapy. Adipose tissue-derived stromal vascular fraction (SVF) comprises a heterogeneous cell population, considered an easily accessible source of MSCs with therapeutic potential in autoimmune diseases. This study aimed to assess the regenerative capacity of low-level laser-activated SVF in an MS cat model. Methods Fifteen adult Persian cats were used in this study: Group I (control negative group, normal cats), Group II (EB-treated group, induced for MS by ethidium bromide (EB) intrathecal injection), and Group III (SVF co-treated group, induced for MS then treated with SVF on day 14 post-induction). The SVF was obtained after digesting the adipose tissue with collagenase type I and injecting it intrathecal through the foramen magnum. Results The results showed that the pelvic limb’s weight-bearing locomotion activity was significantly (P ≤ 0.05) recovered in Group III, and the Basso, Beattie, and Bresnahan (BBB) scores of hindlimb locomotion were significantly higher in Group III (14 ± 0.44) than Group II (4 ± 0.31). The lesion’s extent and intensity were reduced in the magnetic resonance imaging (MRI) of Group III. Besides, the same group showed a significant increase in the expression of neurotrophic factors: BDNF, SDF and NGF (0.61 ± 0.01, 0.51 ± 0.01 and 0.67 ± 0.01, respectively) compared with Group II (0.33 ± 0.01, 0.36 ± 0.006 and 0.2 ± 0.01, respectively). Furthermore, SVF co-treated group revealed a significant (P ≤ 0.05) increase in oligodendrocyte transcription factor (Olig2) and myelin basic protein (4 ± 0.35 and 6 ± 0.45, respectively) that was decreased in group II (1.8 ± 0.22 and 2.9 ± 0.20, respectively). Moreover, group III showed a significant (P ≤ 0.05) reduction in Bax and glial fibrillary acidic protein (4 ± 0.53 and 3.8 ± 0.52, respectively) as compared with group II (10.7 ± 0.49 and 8.7 ± 0.78, respectively). The transmission electron microscopy demonstrated regular more compact, and markedly (P ≤ 0.05) thicker myelin sheaths (mm) in Group III (0.3 ± 0.006) as compared with group II (0.1 ± 0.004). Based on our results, the SVF co-treated group revealed remyelination and regeneration capacity with a reduction in apoptosis and axonal degeneration. Conclusion SVF is considered an easy, valuable, and promising therapeutic approach for treating spinal cord injuries, particularly MS.

Combined laser-activated SVF and PRP remodeled spinal sclerosis via activation of Olig-2, MBP, and neurotrophic factors and inhibition of BAX and GFAP

“…The present study provides evidence that a single treatment involving the combination of laser-activated PRP and stromal vascular fraction in a platelet-rich plasma vehicle can effectively promote functional and structural recovery in spinal cord demyelination injury. This could be declared as cats in the SVF + PRP group exhibited noticeable functional improvement, gait coordination, trunk stability, and higher BBB scores compared to the control positive group as reported in prior research 45 , 46 and 20 , 33 in each treatment alone. The combination of SVF + PRP was found to alleviate the severity and intensity of multiple sclerosis in MRI imaging by reducing swelling, necrosis, and apoptosis, as well as promoting the growth of new neurons and axons better than 20 , 33 which is unlike that mentioned by 47 .…”

“…The tube was incubated at 37 °C for 90 min with continuous agitation. After the digestion period, an equal volume of Dulbecco’s modified Eagle’s medium (DMEM) was added to neutralize the collagenase activity 20 .…”

“…The supernatant was thoroughly aspirated, leaving behind the cell pellets consisting of the stromal vascular fraction. The viability of the cells was assessed using trypan blue dye, and cell counts were performed using a hemocytometer 20 .…”

“…The parameters for these sequences, including the repetition time (TR), echo time (TE), and inversion time (TI), were as follows: TR/TE 2880/111 ms for dorsal T2-weighted, TR/TE 623/1 ms for dorsal T1-weighted, TR/TE 3290/99 ms for transverse T2-weighted, TR/TE 651/12 ms for transverse T1-weighted, and TR/TE/TI 3310/61/140 ms for sagittal STIR. The imaging protocol covered the region from the T11 to L3 vertebral bodies 20 , 33 .…”

“…SVF is a complex mixture of cells, including cells with the stimulatory ability and other stromal cells and signaling molecules. The therapeutic potential of SVF includes spinal cord injury 19 , MS 20 , ischemic heart failure 21 , wound healing 22 , and osteoarthritis 23 . The regenerative properties of SVF, attributed to its composition of ADSCs, have been investigated extensively.…”