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

A single injection of platelet-rich plasma (PRP) or stromal vascular fraction (SVF) in treating neurological ailments suggests promise; however, there is limited evidence of the efficacy of combination therapy. This trial aimed to determine whether combining SVF and PRP could provide further therapeutic effects in treating multiple sclerosis (MS). Fifteen Persian cats were separated into three groups (n = 5): group I (control negative), and group II (control positive); EB was injected intrathecally into the spinal cord and then treated 14 days later with intrathecal phosphate buffered saline injection, and group III (SVF + PRP), cats were injected intrathecally with EB through the spinal cord, followed by a combination of SVF and PRP 14 days after induction. Therapeutic effects were evaluated using the Basso-Beattie-Bresnahan scale throughout the treatment timeline and at the end. Together with morphological, MRI scan, immunohistochemical, transmission electron microscopy, and gene expression investigations. The results demonstrated that combining SVF and PRP successfully reduced lesion intensity on gross inspection and MRI. In addition to increased immunoreactivity to Olig2 and MBP and decreased immunoreactivity to Bax and GFAP, there was a significant improvement in BBB scores and an increase in neurotrophic factor (BDNF, NGF, and SDF) expression when compared to the positive control group. Finally, intrathecal SVF + PRP is the most promising and safe therapy for multiple sclerosis, resulting in clinical advantages such as functional recovery, MRI enhancement, and axonal remyelination.

Anatomical, histological, and histochemical alterations in portio vaginalis uteri with an evaluation of the vaginal artery vascularity during the luteal and early pregnant stages in domestic buffaloes (Bubalus bubalis).

The portio-vaginalis uteri (PVU) and its mucus secretion have shown an essential role in conception. A significant endeavour to improve buffaloes' reproductive efficiency is the investigation of their basic reproductive pattern, which provides a reference for applications in breeding and pregnancy. The present study aimed to evaluate the anatomical and histological alterations in PVU regarding to the vaginal artery (VA) hemodynamic at luteal and early pregnant stages in buffalos. Egyptian live buffaloes (n = 16) and fresh genitals (n = 25) of mature buffalo were used. Different luteal and early pregnant stages were macroscopically identified with the shape and mucosal colouration with discharges of the PVU. Histological examination showed a significant difference in area % of alcian blue and periodic acid Schiff positive granules which considered an indication for presence of acidic and neutral mucins respectively in the epithelial cells of PVU mucosa which increased in pregnant stage than in other luteal stages. VA assessment demonstrated an increase in luminal diameter and thickness of tunica muscularis in pregnant stage than other stages (P < 0.05). Middle uterine (MUA) and VA arteries peak velocity point (PSV mm/sec) were elevated (P < 0.05) in pregnant stage, with a marked reduction in both resistance and pulsatility indices (RI and PI), and ratio of systolic /diastolic (S/D). Positive correlation was detected between VA. PSV and, MUA. PSV (r = 0.87), but a negative relation was detected with VA. S/D (r = -0.77), VA.PI (r = -0.89), VA. RI (r = -0.97), MUA. S/D (r = -0.94), MUA. PI (r = -0.85), and MUA. RI (r = -0.88). Doppler indices were negatively corrected with the VA. PSV (r = -0.68). It was concluded that there was a significant alterations in histological features of the cervical PVU at different physiological stages (luteal and early pregnant) in buffalos in relation to the MUA and VA hemodynamic pattern and that hypotheses can be established regarding the female cyclicity that affected by both arteries hemodynamics change.

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

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.

A novel cell-free intrathecal approach with PRP for the treatment of spinal cord multiple sclerosis in cats.

BACKGROUND: Multiple sclerosis (MS) is a progressive autoimmune demyelinating disease of the central nervous system. To date, there is no effective therapy for it. Our study aimed to determine the potential role of platelet-rich plasma (PRP) in the treatment of MS in cats. METHODS: The current study was conducted on 15 adult Persian cats that were divided into three groups: control negative, control positive (ethidium bromide (EB)-treated group), and PRP co-treated group (EB-treated group intrathecally injected with PRP on day 14 post-spinal cord injury). PRP was obtained by centrifuging blood on anticoagulant citrate dextrose and activating it with red and green laser diodes. The Basso-Beattie-Bresnahan (BBB) scores were used to assess the motor function recovery on days 1, 3, 7, 14, 20, and 28 following 14 days from EB injection. Moreover, magnetic resonance imaging (MRI) analysis, histopathological investigations, transmission electron microscopy (TEM) studies, and immunohistochemical analysis were conducted, and the gene expressions of nerve growth factors (NGFs), brain-derived neurotrophic factors (BDNF), and stromal cell-derived factors (SDF) were evaluated. RESULTS: Our results indicated that PRP had a significant ameliorative effect on the motor function of the hindlimbs as early as day 20 and so on. MRI revealed that the size and intensity of the lesion were significantly reduced in the PRP co-treated group. The histopathological and TEM investigations demonstrated that the PRP co-treated group had a significant improvement in the structure and organization of the white matter, as well as a high remyelination capacity. Furthermore, a significant increase in myelin basic protein and Olig2 immunoreactivity as well as a reduction in Bax and glial fibrillar acidic protein immune markers was observed. NGFs were found to be upregulated by gene expression. CONCLUSION: As a result, we concluded that the intrathecal injection of PRP was an effective, safe, and promising method for the treatment of MS.

Histological and hemodynamic characterization of corpus luteum throughout the luteal phase in pregnant and non-pregnant buffalos in relation to nitric oxide levels based on its anatomical determination.

This study aims to compare the complete growth and development of corpus luteum (CL) in domestic buffalos from day 5 until day 40 after ovulation either in pregnant or non-pregnant animals and whether luteal vascularity (LV) with progesterone (P4) and nitric oxide (NO) could determine luteal functionality or not. Pluriparous buffalos (Bubalus bubalis) were categorized as pregnant (n = 6) or non-pregnant (n = 9) after pregnancy check at day 25. Animals were subjected to ultrasound analysis to determine the CL area (cm2) and LV. Blood sampling was performed following the Doppler examination. Ovarian tissue samples from non-pregnant buffalo genitalia (n = 18) and early pregnant buffalo genitalia (n = 3) were collected from great abattoirs. Luteal Doppler indices were lower in the pregnant group, while peak systolic velocity (PSV) was increased (p < 0.05) in the same pregnant females. Both P4 and NOMs were elevated (p < 0.05) in the pregnant group. There was a positive correlation (p < 0.01) between P4 and CL PSV. Based on our macroscopical examination, the CL of non-pregnant buffalos was classified into four stages. Histologically, stage I showed that CL was covered by a highly vascularized connective tissue (CT) capsule. It consisted of small and large lutein cells, whereas stage II was similar to stage I except for the presence of numerous fibroblast cells and vacuolated cells. Stage III was characterized by increasing the number of collagen fibers and the thickness of the blood vessels. Stage IV revealed thickening of the CT capsule and septae, regressed capillaries and arterioles, in addition to shrunken degenerated lutein cells. CL of pregnant buffalos revealed the same structure as CL at stage II. CL area was increased in the pregnant group. The collective data suggested that evaluation of the luteal artery could be extremely helpful to determine the potential benefits of colored and pulsed Doppler in CL vascularization assessment in both luteal and early pregnancy phases.

Ovarian, uterine, and luteal vascular perfusions during follicular and luteal phases in the adult cyclic female rabbits with special orientation to their histological detection of hormone receptor.

Understanding the does reproductive hemodynamic changes during the estrous cycle is crucial for improving reproductive competence and fertility potential in this species. The objective of this study is to investigate the hemodynamic variations in ovarian (OA) and uterine (UA) arteries, histological and morphometric changes in ovarian and uterine tissues throughout the follicular (FP) and luteal (LP) phases in rabbits and determine estrogen (ER), progesterone (PR) receptors, and vascular endothelial growth factor (VEGF) distributions using immunohistochemistry.Fourteen adults pluriparous New Zealand rabbits were divided into rabbits at the FP (Day - 1; n = 7) and those at the LP (Day 9; n = 7). Animals were subjected to Doppler, hormonal (estrogen [E2], progesterone [P4], insulin-like growth factor [ILGF], and VEGF), histological, and immunohistochemical analyses. In LP, OA Doppler indices were significantly increased, whereas peak systolic velocity (PSV) was decreased compared with that in FP. UA Doppler indices were significantly decreased in the LP, whereas PSV was increased (P < 0.05). E2 levels were increased in the FP, whereas P4 levels were increased in the LP. The morphometric analysis of uterine tissues during the LP revealed an increase in the mean uterine endometrium length, endometrial connective tissue area percentage (%), endometrial glands number, myometrial area (%) and thickness. Furthermore, ovarian follicles and corpus luteum (CL) displayed strong positive immunoreactivity for ER, PR, and VEGF-A during both phases. The ovarian sections displayed a substantial (P < 0.05) increase in the area % of VEGF-A in the ovarian follicles during FP while in the CL during LP. Conversely, area percentage of VEGF-A immunoreactivity in the uterine luminal and glandular epithelia during the FP and LP revealed no differences. However, the number of VEGF-A-stained blood capillaries revealed an increase during LP than FP. In conclusion, this study demonstrated for the first time the changes in both ovarian and uterine arteries during two different phases of the rabbit cycle in relation to the histo-morphometric analysis and distribution of ER, PR, and VEGF-A, which regulate uterine functions that play a role in reproduction.

Ovarian, uterine and luteal hemodynamic variations between pregnant and non-pregnant pluriparous Egyptian buffalos with special reference to their anatomical and histological features.

This study determined the ovarian, uterine, and luteal hemodynamic variations using Doppler between pregnant and non-pregnant pluriparous buffalos in relation to their anatomical and histological basics during the first 31 days after natural mating. Adult healthy cyclic Egyptian buffalo (n = 10) were selected and categorized into two groups; group 1 (n = 5) was mated naturally by a fertile bull during the late estrus phase, and group 2 (n = 5) was not mated. Animals were subjected to Doppler ultrasonography to evaluate luteal, ovarian, and uterine blood flows from day 7 until day 31 post-mating. Besides, three pregnant (one month) and other non-pregnant uterus (n = 6) were obtained from a local abattoir to study the anatomical and histological features. Our results revealed that the luteal, ovarian, and uterine arteries cross-sectional diameters/mm increased (P < 0.05) from day 7 till day 31. Resistance (RI) and pulsatility indices (PI) decreased linearly (P < 0.05) in pregnant buffalos till day 31, but the peak systolic, end diastolic velocities and flow volume of those arteries were increased. Additionally, luteal colored areas away and toward CL were increased (P < 0.05) in the pregnant group compared to non-pregnant ones. There was a significant (P < 0.05) increase in the lumen diameter of luteal, ovarian, and uterine artery sections in pregnant buffalos compared to those of non-pregnant ones. While the mean value of tunica media's thickness of both luteal and uterine artery was significantly higher in non-pregnant buffalos than pregnant ones, except for that of the ovarian artery. Additionally, the ovarian and uterine artery tunica muscularis relative area % was (P < 0.05) higher in pregnant buffalos than in non-pregnant ones, except for that of the luteal artery. It was concluded that in pregnant buffalos, ovarian, uterine, and luteal blood flows were improved from the first week until 31 days post-mating via a decline in both Doppler indices with an increase in Doppler velocities and blood flow volume in relation to their histological changes based on their anatomical architecture in comparison to non-pregnant one.