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

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          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.

          Supplementary Information

          The online version contains supplementary material available at 10.1186/s13287-022-03222-2.

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          Most cited references96

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            PUVA-induced repigmentation of vitiligo: scanning electron microscopy of hair follicles.

            PUVA-i-duced repigmentation of vitiligo was studied using both the split-dopa reaction and scanning electron microscopy. Proliferation of hypertrophic, Dopa-positive melanocytes were observed in the lower portion of some hair follicles, whereas other giant melanocytes were observed along the middle portion. The existence of a melanocyte reservoir in human hair follicles is postulated.
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              A sensitive and reliable locomotor rating scale for open field testing in rats.

              Behavioral assessment after spinal cord contusion has long focused on open field locomotion using modifications of a rating scale developed by Tarlov and Klinger (1954). However, on-going modifications by several groups have made interlaboratory comparison of locomotor outcome measures difficult. The purpose of the present study was to develop an efficient, expanded, and unambiguous locomotor rating scale to standardize locomotor outcome measures across laboratories. Adult rats (n = 85) were contused at T7-9 cord level with an electromagnetic or weight drop device. Locomotor behavior was evaluated before injury, on the first or second postoperative day, and then for up to 10 weeks. Scoring categories and attributes were identified, operationally defined, and ranked based on the observed sequence of locomotor recovery patterns. These categories formed the Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale. The data indicate that the BBB scale is a valid and predictive measure of locomotor recovery able to distinguish behavioral outcomes due to different injuries and to predict anatomical alterations at the lesion center. Interrater reliability tests indicate that examiners with widely varying behavioral testing experience can apply the scale consistently and obtain similar scores. The BBB Locomotor Rating Scale offers investigators a more discriminating measure of behavioral outcome to evaluate treatments after spinal cord injury.
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                Author and article information

                Contributors
                mariamfayez45@yahoo.com
                yarasayed89@gmail.com , yarasayed89@cu.edu.eg
                nohayassin428@yahoo.com , nohayassin428@cu.edu.eg
                Asmaa.khairy@cu.edu.eg
                proteome@cu.edu.eg
                Marwa199@gmail.com , Marwaibrahim@cu.edu.eg
                hamdyrezk81@cu.edu.eg
                Journal
                Stem Cell Res Ther
                Stem Cell Res Ther
                Stem Cell Research & Therapy
                BioMed Central (London )
                1757-6512
                11 January 2023
                11 January 2023
                2023
                : 14
                : 6
                Affiliations
                [1 ]GRID grid.7776.1, ISNI 0000 0004 0639 9286, Department of Anatomy and Embryology, Faculty of Veterinary Medicine, , Cairo University, ; Giza, 12211 Egypt
                [2 ]GRID grid.7776.1, ISNI 0000 0004 0639 9286, Department of Cytology and Histology, Faculty of Veterinary Medicine, , Cairo University, ; Giza, Egypt
                [3 ]GRID grid.7776.1, ISNI 0000 0004 0639 9286, Department of Pathology, Faculty of Veterinary Medicine, , Cairo University, ; Giza, Egypt
                [4 ]GRID grid.7776.1, ISNI 0000 0004 0639 9286, Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, , Cairo University, ; Giza, Egypt
                [5 ]GRID grid.511523.1, ISNI 0000 0004 7532 2290, Department of Biomedical Research, , Armed Forces College of Medicine, ; Cairo, 12211 Egypt
                Author information
                http://orcid.org/0000-0003-0802-3648
                http://orcid.org/0000-0003-1468-8515
                http://orcid.org/0000-0001-8985-2207
                http://orcid.org/0000-0002-6155-8871
                http://orcid.org/0000-0001-6305-9913
                Article
                3222
                10.1186/s13287-022-03222-2
                9832640
                36627662
                9e8653d8-d013-46e8-9a8d-c94ae203869c
                © The Author(s) 2022

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                : 28 July 2022
                : 14 December 2022
                Funding
                Funded by: Cairo University
                Categories
                Research
                Custom metadata
                © The Author(s) 2023

                Molecular medicine
                multiple sclerosis,stromal vascular fraction,mesenchymal stem cells,flow cytometry,mri,immunohistochemistry,tem

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