The Role of Stem Cells Derived From the Mesenchyme of the Umbilical Cord in Reducing Immunosuppressive Drug Doses Used in Allogenic Transplantations.

BACKGROUND: This study evaluated the potential of Wharton's jelly mesenchymal stem cells with high tolerogenic properties in reducing immunosuppressive dosage and related adverse effects. METHODS: A 4- to 6-week-old, 30-40 g weight, male inbred CD57BL/6 mice were used as skin allograft donors, whereas Balb/c mice with similar characteristics were used as recipients. Wharton's jelly stem cells were obtained from a commercial kit sourced from human umbilical cord. Skin allografts were performed from CD57Bl6 to Balb/c mice (day 0). Group 1 (control) received no treatment. Group 2 received 15 mg/kg cyclosporin A on days 0 to 30. Group 3 received 5.7 × 10 6 and 10.3 × 10 6 cell/kg Wharton's jelly stem cells on days 0 and 3, respectively. Groups 4, 5, and 6 received a combination of 15, 10, and 5 mg/kg per day cyclosporine A (days 0 to 30) with the same stem cell dose with group 3, respectively. Graft rejection was evaluated with digital photography and thermal imaging, histopathology (Banff grading, epithelialization scores, dermoepidermal dissociation), immunochemistry (Ki-67 and Bcl-2), and biochemical methods (interleukin 10, interleukin 2, interferon γ, tumor necrosis factor α) (day 10). Cumulative adverse effects of cyclosporin A occurring in the groups were revealed by histopathological evaluation of kidney and liver (a modified semiquantitative method of infiltration of inflammatory cells around the portal area and lobular region in liver; modification of the Banff rating of proximal tubules and hypertrophia of juxtaglomerular apparatus cells in kidney) (day 30). RESULTS: There was no rejection in groups 2, 4, and 5 until the end of study. These were statistically different versus groups 1 (day 10 ± 0.71), 3 (day 11 ± 0.82), and 6 (day 11 ± 0.58) (all P 's < 0.05). Groups 4 and 5 have exhibited statistically similar findings in histopathological (4 epithelization score: 3.7 ± 1.3; 5 epithelization score: 3.5 ± 0.5; 4 Banff grading score: 0.8 ± 0.6; 5 Banff grading score: 1.0 ± 0.5; both P 's = 1.00), immunohistochemical (4 Bcl-2 score: 3.5 ± 0.5, P = 0.618; 5 Bcl-2 score: 3.4 ± 0.5, P = 1.00; 4 Ki-67 score: 3.7 ± 0.4, P = 1.00; 5 Ki-67 score: 3.5 ± 0.5, both P 's = 1.00), and levels of cytokines (both P 's = 1.00) versus group 2. Adverse effects on kidneys and liver were lowest and statistically similar in groups 3, 5, and 6 (all P 's = 00) versus group 1. CONCLUSIONS: Wharton's jelly mesenchymal stem cells alter bioavailability of cyclosporine, albeit at much lower doses and with fewer systemic adverse effects.

Effects of Adipose-Derived Stem Cells and Platelet-Rich Plasma for Prevention of Alopecia and Other Skin Complications of Radiotherapy.

BACKGROUND: Radiotherapy (RT) involves the use of ionizing radiation in treating malignancies and benign disorders. However, RT damages target and healthy surrounding tissues in a dose-dependent manner. This effectively reduces patient compliance and quality of life, thereby warranting the prevention of RT-induced adverse effects on skin. Adipose-derived stem cells (ASCs) are used to treat RT-induced damage and platelet-rich plasma (PRP) provides a scaffold that potentiates the effects of ASCs. Thus, the aim of this study was to determine the mechanism employed by ASCs and PRP in protecting against RT-induced adverse effects. METHODS: We have established an immunodeficient mouse transplantation model using which human hair follicular units were implanted. When the follicular units were macroscopically and microscopically mature and anagenic, we administered localized RT. Subsequently, the mice were randomly divided into 4 groups based on the subcutaneous injection of the following to the irradiated transplantation site: saline, PRP, ASCs, and a combination of ASCs and PRP. Next, we used macroscopic and microscopic analyses to determine the protective effects of the injected solutions on skin and hair follicles. RESULTS: Adipose-derived stem cells reduced RT-induced adverse effects, such as impaired wound healing, alopecia, skin atrophy, and fibrosis by suppressing inflammation, dystrophy, degeneration, connective tissue synthesis, and apoptosis and increasing cellular proliferation, differentiation, and signaling. Moreover, these effects were augmented by PRP. CONCLUSIONS: Thus, co-administering ASCs with PRP in mice prevented RT-induced adverse effects and can be tested for use in clinical practice.

Investigation of the Effect of Hafnium Chloride on Sperm Viability and Motility in Normospermic Cases: An In Vitro Study.

INTRODUCTION: Hafnium alloys are employed in medical applications due to their biocompatibility and high corrosion resistance. These alloys have demonstrated osteogenic and antimicrobial activities in surgical implants and have been utilized in the treatment of sarcoma. Additionally, a sensor based on hafnium nanoparticles has been reported for the detection of coronavirus disease 2019. Despite the increasing usage of hafnium, a literature review reveals no studies examining its effects on sperm in both human and animal species. METHODS: Semen samples were analyzed according to the 2010 World Health Organization (WHO) criteria, and 20 normospermic specimens were included in the study. Three groups were formed: control, hafnium chloride 2 mg/mL, and 4 mg/mL. Motility and viability were assessed in all groups at the 20th and 40th minutes. RESULTS: The decrease in viable sperm count was found to be significant in the 2 mg/ml HfCl4 group (difference: 12.73 ± 0.8, p<0.001) and the 4 mg/ml HfCl4 group (difference: 41.72 ± 1.34, p<0.001) compared to the control group. A time-dependent decrease in sperm viability was significant across all groups (difference: 8.93 ± 0.59, p<0.001). The decrease in viable sperm count in the 4 mg/ml HfCl4 group was significant when compared to the 2 mg/ml HfCl4 group (difference: 29 ± 1.27, p<0.001). The decrease in total motile sperm count was observed in both the 2 mg/ml HfCl4 group (difference: 12.80 ± 1.30, p<0.001) and the 4 mg/ml HfCl4 group (difference: 35.63 ± 1.12, p<0.001) compared to the control group. Additionally, the decrease in total motile sperm count in the 4 mg/ml HfCl4 group was significant compared to the 2 mg/ml HfCl4 group (difference: 22.80 ± 1.60, p<0.001). A time-dependent decrease in total motile sperm count was also significant (difference: 6.03 ± 0.49, p<0.001). CONCLUSION: The study determined that hafnium chloride negatively affects sperm motility and viability in vitro. These effects may be due to the presence of an acidic environment. It has been demonstrated that instruments containing this element may pose a potential risk.

Histopathological Evaluation of Platelet-Rich Plasma Effect in Acute Tympanic Membrane Perforation.

Platelet-rich plasma (PRP) is a reliable and has low side-effect profile and has beneficial effects on wound healing. Its investigatory effects on wound-healing process were shown on various tissues. This study aims to investigate PRP's local application effects to perforated rat TM in terms of healing and histopatological outcomes. Twenty-two Wistar rats were used in the study. The rats' ears were examined with a pediatric endoscope (2.7 mm, 0°), and the TM posterior quadrant of their right ear was perforated with a 20-gauge needle. After this procedure, the rats were divided into two equal groups. A spongel with PRP was applied on the perforated TM in the first group, and spongel with standard saline solution was applied on the second group. Following the sacrifice, the middle air bullas were carefully dissected and removed for histopathological examination. Hematoxylin eosin (for fibroblasts, lymphocyte, collagen fibers) and immunohistochemical staining were done for epithelial growth factor receptor (EGFR), fibroblast growth factor receptor (FGFR1), and vascular endothelial growth factor (VEGF) staining for histopathologic examinations. There was not a significant difference between the two groups for lymphocyte. There was a significant difference between control and study groups for collagen and EGFR (P < 0.05). Although the mean value of FGF- and VEGF-positive cells was higher in the study group than in the control group, the difference was not significant (P > 0.05). PRP is an effective autologous material for the healing process of acute TM perforations in a rat model, as demonstrated in the present study. We think that the use of PRP for acute TM perforations can have a positive effect on the healing process by increasing the level of growth factors, especially EGFR. In addition, an increase in collagen can also have a positive effect on healing. Supplementary Information: The online version contains supplementary material available at 10.1007/s12070-021-02912-2.

Treatment of Acute Vocal Fold Injury With Platelet-Rich Plasma.

OBJECTIVES: Platelet-rich plasma (PRP) is a reliable and has low side-effect profile and has beneficial effects on wound healing. Its investigatory effects on wound-healing process were shown on various tissues. The aim of the present study was to evaluate effectiveness of PRP application on scar tissue of acute vocal fold injury. MATERIALS AND METHODS: Twenty-four Wistar rats were used in the study. The entire layer of the lamina propria down to the thyroarytenoid muscle of 10 subjects was unilaterally injured by with a microscissor. Gelfoam-absorbed PRP was applied on the injured area for 10 minutes. Control group consisted of rats unilaterally injured using a microscissor, and gelfoam with normal saline was applied on the injured area. Following sacrifice, the larynxes were carefully dissected and removed for histopathologic examination. After excised larynx experiments, serial sections were prepared from vocal fold. Hematoxylin eosin and immunohistochemical staining were done for epithelial growth factor receptor (EGFR), fibroblast growth factor receptor (FGFR1), and vascular endothelial growth factor (VEGF) staining for histopathologic examinations. RESULTS: There was not a significant difference between the two groups for lymphocyte. Although collagen and VEGF were higher in the study group, there was not a significant difference between the groups (P > 0.05). There was a significant difference between control and study groups for EGFR and FGFR1(P < 0.05). CONCLUSIONS: PRP has beneficial effects on wound healing. PRP accelerates epithelization of injured rat vocal folds by inducing EGFR secretion. PRP is an autogenous, reliable, low side-effect profile, easily harvested material. PRP may be useful to prevent scar formation.

Intra-articular platelet-rich plasma injection for the treatment of temporomandibular disorders and a comparison with arthrocentesis.

BACKGROUND: Temporomandibular joint (TMJ) internal derangements are progressive painful conditions and cause joint dysfunction, joint sound, malocclusion, and locking of the mouth. Conservative and invasive techniques can be used for the treatment of TMJ internal derangements. The objective of the present study was to examine the benefit of an intra-articular platelet-rich plasma (PRP) injection and to compare this with arthrocentesis. METHODS: Twenty patients (female: male; 15:5; age 26, 3 ± 9.3 years) for a total of 32 joints with reducible anterior disc dislocation, as confirmed by Magnetic Resonance Imaging (MRI), were divided into two groups. PRP was used for the study group, and arthrocentesis was used for the control group. Pain intensity, maximal interincisal opening, and TMJ sounds were assessed and compared for evaluation of treatment success. RESULTS: There was a statistically significant reduction in pain intensity and joint sound and an increase in mouth opening in the study group when compared with the control group. CONCLUSIONS: This study shows that intra-articular PRP injection for the treatment of reducible disc displacement of the TMJ is a more effective method than arthrocentesis.

Improvement in embryo quality and pregnancy rates by using autologous cumulus body during icsi cycles.

OBJECTIVE: To determine whether the addition of intact cumulus cell mass (ICM) to both embryo culture (EC) and embryo transfer (ET) improves embryo quality and pregnancy rates. MATERIAL AND METHODS: A total of 133 infertile couples were included, of which 67 received ICM (study group) and 66 did not (control group). The ICM was obtained from a simple cutting of the cumulus corona oocyte complex (CCOC). A case control study design was used. RESULTS: The clinical characteristics of the two groups before the embryo culturing step were similar with respect to age, estradiol level on the day of hCG and endometrial thickness on the day of embryo transfer (p>0.05). On the other hand study group with ICM had higher number of high quality embryos (3.1±1.4 vs 2.4±1.1, p=0.03), higher implantation rate (53.7% vs 34.8%, p=0.02) and higher ultrasound confirmation of gestational sac and fetal heart beat as ongoing pregnancy rates (44.7% vs 27.2%, p=0.04) compared to the control group without ICM. CONCLUSION: Addition of ICM improves embryo quality and pregnancy rates. This is a cost-and time-effective simple procedure that shows great promise for the improvement of infertility treatment.

Effects of Lipokit® centrifugation on morphology and resident cells of adipose tissue / Efectos de la centrifugación de Lipokit® sobre la morfología y las células residentes del tejido adiposo

The aim of adipose tissue engineering is creating autologus vascularized fat tissue to be used for practical soft tissue reconstruction in human clinic. Unfortunately, in practice, long-term results of fat transplantation are often untrustworthy and unreliable, to overcome this problem different many lipoinjection techniques developed in the last 20 years. Centrifuge is a fundamental stepin the preparation of adipose tissue. We focused on some cell markers especially MSCs markers and histological structural properties after with lipokit centrifugation and without lipokit centrifugation of adipose tissue obtained by liposuction by this new technique. Adipose tissue was taken by liposuction and separates to two portions. One of them is centrifugated by Lipokit machine (C+) has a micro filter and the other is not (C-). After centrifugation smear slides and paraffin sections were prepared from these tissues. These slides were stained with H&E and Toluidine Blue. Paraffin sections were immunohistochemically stained with CD34, von Willebrand Factor, CD73, CD90 and CD105. Smear preparations showed a continuous three dimensional plasma membrane appearance of adipocytes. C+ and C- showed expression of CD34, von Willebrand Factor, CD73, CD90, CD105. C+ seems to have more free cells expressing than C-. While passing the filter of Lipokit, large adipocytes and connective tissue parts disintegrate and thus increases the surface area of lipoaspirate. Lipokit® machine release the group cells which are necessary for angiogenesis and they become more freely to construct angiogenesis.

El objetivo de la ingeniería del tejido adiposo es la creación de tejido graso vascularizado autólogo para ser utilizado en clínica humana para la reconstrucción de tejido blando. Desafortunadamente en la práctica, los resultados a largo plazo del trasplante de grasa son poco fiables y no seguros; para superar este problema, se han desarrollado en diferentes países, en los últimos 20 años, variadas técnicas de lipoinyección. La centrifugación es un paso fundamental en la preparación del tejido adiposo. Nos hemos centrado en algunos marcadores, especialmente, de células precursoras mesenquimales y propiedades histológicas estructurales después de la centrifugación mediante Lipokit y sin la centrifugación por Lipokit del tejido adiposo obtenido mediante liposucción. El tejido adiposo fue tomado por liposucción y se separó en dos porciones. Una se centrifugó mediante el sistema Lipokit (C +), con un microfiltro y la otro no (C-). Después de centrifugación, muestras del frotis y secciones de parafina se prepararon a partir de estos tejidos. Los frotis se tiñeron con H&E y azul de toluidina. Las secciones de parafina se tiñeron inmunohistoquímicamente con CD34, factor de von Willebrand, CD73, CD90 y CD105. Las preparaciones de los frotis mostraron una apariencia tridimensional continua de la membrana plasmática de los adipocitos. Tanto en C+ y C- se observó la expresión de CD34, factor de von Willebrand, CD73, CD90 y CD105. En C+ parecen expresarse más células libres que en C-. Cuando se utilizó el filtro de Lipokit, los adipocitos grandes y partes del tejido conectivo se desintegraron, por lo tanto aumentó el área de superficie de lipoaspirado. El sistema Lipokit® libera los grupos celulares que son necesarios para la angiogénesis y se hacen más libres para promoverla.

Assessment of Spermatozoa Morphology under Light Microscopy with Different Histologic Stains and Comparison of Morphometric Measurements / Evaluación de la Morfología de los Espermatozoides bajo Microscopía Óptica con Diferentes Tinciones Histológicas y Comparación de las Mediciones Morfométricas

The aim was to examine the morphology of spermatozoa with different staining methods and aimed to find the better staining methods for morphology of spermatozoa in our study. Randomized 67 patients taken for the study who were admitted to Assisted Reproductive Techniques Unit. In the first part of the study, smears were stained with Hematoxylin Eosin (HE), Toluidin Blue (TB), Giemsa, Wright, ferrous Weigert haematoxylin stain, Orange G, eosin-aniline blue dye, Shorr Method, Papanicolau, Berg Method, Light Green stain, Acridine Orange (AO) and Janus Green dyes. In the second part of the study, smear preparations of 10 patients with normozoospermic were stained with HE, Toluidin Blue (TB), Shorr Method and Papanicolau. Four measurements were made including the middle piece, head length- head width and tail length for 200 spermatozoa with normal morphology. Comparisons were made between the stains that which showed a better morphology. Condensation assessment was not possible in smears stained with Shorr, Berg, Method, AO. Better assessment of condensation could be made in other stains. In the second part the smallestvalues belonged to of TB stain according to measurements of head of the spermatozoa. There was a significant difference at the head length with TB stain. Although measurements of Shorr and Papanicolau areclose to each otherand the largestvalues belonged to Papanicolau dye. It was concluded that measurement values in human sperm morphology could alter with the used staining method.

El objetivo fue examinar la morfología de los espermatozoides con diferentes métodos de tinción y encontrar los mejores métodos para su estudio. Fueron seleccionados para el estudio, de manera aleatoria 67 pacientes, quienes ingresaron a la Unidad de técnicas de reproducción asistida. En la primera parte del estudio, se realizaron y tiñeron frotis con hematoxilina eosina (HE), Azul de Toluidina (AT), Giemsa, tinción de Wright, Hematoxilina Férrica de Weigert, Anaranjado G, tinción eosina-anilina, método de Shorr, Papanicolau, método Berg, tinción verde brillante, anaranjado de acridina (AO) y tinción verde Janus. En la segunda parte del estudio, se realizaron frotis de 10 pacientes con normozoospérmicos y se tiñeron con HE, AT, Método Shorr y Papanicolau. Se realizaron cuatro mediciones: ancho de la cabeza, longitud de la cabeza, parte media y cola, sobre 200 espermatozoides con morfología normal. Se compararon las tinciones que mostraban mejor la morfología. La evaluación de la compactación no fue posible en los frotis teñidos con los métodos de Shorr, Berg y AO. Una mejor evaluación de la copactación podría hacerse en otras tinciones. En la segunda parte los valores menores correspondieron a la tinción de AT en relación a la medición de la cabeza de los espermatozoides. Hubo una diferencia significativa en la longitud de la cabeza con tinción de AT. Las mediciones en los frotis con técnicas de Shorr y Papanicolau fueron similares, con valores más altos bajo tinción de Papanicolau. Se concluyó que los valores de la medición morfológica en espermatozoides humanos podrían ser alterados según el método de tinción utilizado.