Saving zone of stasis in burn wounds by nanoliposomal Mg-ATP.

ATP is a crucial molecule for every energy-dependent process in cells. In ischemic tissues, ATP production declines, and it finally results in cell death. One of the most common strategies in burn wound management is saving the zone of ischemia. In the current study, Mg-ATP-containing nanoliposomes were formulated and studied in vitro and in vivo. The particle size of the vesicles was between 50 and 100 nm and the mean zeta potential was -4.05 ± 0.52 mV as evaluated by dynamic light scattering and Zeta sizer instrument, respectively. The encapsulation efficiency of ATP in the nanoliposomes was found to be 9.3%. The morphology and size of nanoliposomes were further studied by transmission electron microscopy. The standard MTT assay revealed no cytotoxicity of the nanoliposomes when tested on the rat fibroblast cells. Forty rats were randomly divided into four groups (N = 10 each). Burn wounds were created by burn comb model on the back of the rats and the zone of stasis in each group was treated every 12 h for 3 days by injecting them with the Mg-ATP-nanoliposomes. Control samples included empty nanoliposomes, unencapsulated Mg-ATP and the Krebs-Henseleit buffer. Laser Doppler flowmetry results revealed that blood perfusion in the zone of ischemia in rats treated with Mg-ATP-nanoliposomes was more than in the other groups (p < 0.05). Histopathology revealed saving zone of stasis by Mg-ATP-nanoliposomes. Findings obtained in this study demonstrated that the formulated Mg-ATP-nanoliposome has the potential to save the stasis zone in burn wounds.

Comparison of osteogenic differentiation potential of induced pluripotent stem cells and buccal fat pad stem cells on 3D-printed HA/ß-TCP collagen-coated scaffolds.

Production of a 3D bone construct with high-yield differentiated cells using an appropriate cell source provides a reliable strategy for different purposes such as therapeutic screening of the drugs. Although adult stem cells can be a good source, their application is limited due to invasive procedure of their isolation and low yield of differentiation. Patient-specific human-induced pluripotent stem cells (hiPSCs) can be an alternative due to their long-term self-renewal capacity and pluripotency after several passages, resolving the requirement of a large number of progenitor cells. In this study, a new biphasic 3D-printed collagen-coated HA/ß-TCP scaffold was fabricated to provide a 3D environment for the cells. The fabricated scaffolds were characterized by the 3D laser scanning digital microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and mechanical test. Then, the osteogenesis potential of the hiPSC-seeded scaffolds was investigated compared to the buccal fat pad stem cell (BFPSC)-seeded scaffolds through in vitro and in vivo studies. In vitro results demonstrated up-regulated expressions of osteogenesis-related genes of RUNX2, ALP, BMP2, and COL1 compared to the BFPSC-seeded scaffolds. In vivo results on calvarial defects in the rats confirmed a higher bone formation in the hiPSC-seeded scaffolds compared to the BFPSC-seeded groups. The immunofluorescence assay also showed higher expression levels of collagen I and osteocalcin proteins in the hiPSC-seeded scaffolds. It can be concluded that using the hiPSC-seeded scaffolds can lead to a high yield of osteogenesis, and the hiPSCs can be used as a superior stem cell source compared to BFPSCs for bone-like construct bioengineering.

Effects of Platelet-Rich Fibrin/Collagen Membrane on Sciatic Nerve Regeneration.

ABSTRACT: Alternative treatment approaches to improve the regeneration ability of damaged peripheral nerves are currently under investigation. The aim of the current study was to evaluate the effects of leucocyte/platelet-rich fibrin (L-PRF) with or without a collagen membrane as a supporter on crushed sciatic nerve healing in a rat model. Recovery of motor function and electrophysiologic measurements were evaluated at 4 weeks postoperatively. The whole number of myelinated axons, peripheral nerve axon density, average nerve fiber diameter (µm), and G-ratio were analyzed and compered among the groups. Functional, electrophysiological, and histological evaluations showed no significant difference among the groups with the exception of the L-PRF with collagen membrane groups that showed relatively positive effects on the functional and histological nerve recovery. In addition, the collagen membrane with L-PRF can be effect in nerve regeneration.

Immediate implant placement following 1-year treatment with oral versus intravenous bisphosphonates: a histomorphometric canine study on peri-implant bone.

OBJECTIVES: Bisphosphonates (BPs) are anti-resorptive medications with inhibitory effects on osteoclasts which decrease bone turnover. The present study aimed to assess the early effects of BPs on peri-implant bone. MATERIALS AND METHODS: Twelve female mongrel dogs were assigned to one control and two experimental groups as follows: 12 months of oral alendronate (ALN, 3.5 mg/kg/week) or intravenous (IV) pamidronate (PAM, 1 mg/kg/week) for experimental groups. Following 8 weeks after implant insertion, the specimens were sacrificed, histological evaluation was performed, and C-terminal telopeptide of collagen I (CTx) analysis was performed on serums. RESULTS: PAM demonstrated the greatest marginal bone level (MBL), 2.64 ± 0.48, followed by ALN and control, 2.5 ± 0.4 and 0.66 ± 0.5 respectively. The least bone-implant contact was observed in PAM group (%39 ± 0.03) while it was significantly greater in the control group (p < 0.05). PAM and ALN demonstrated < 10 CTx levels versus > 300 in controls. CONCLUSION: It is assumed that long-term oral or IV BP therapy decreases marginal bone resorption and osseointegration. These changes were more accentuated in IV BP administration. CTx does not seem to be a precise predictor for bisphosphonate-related osteonecrosis of the jaw. CLINICAL RELEVANCE: BPs impair peri-implant bone remodeling and this phenomena may be effective for reducing resorption at esthetic zone implants; however, decrease in osseointegration may be a concern for implant treatment in patients receiving BPs.

Intrapulmonary autologous transplant of bone marrow-derived mesenchymal stromal cells improves lipopolysaccharide-induced acute respiratory distress syndrome in rabbit.

BACKGROUND: Lung diseases such as acute respiratory distress syndrome (ARDS) have a high incidence worldwide. The current drug therapies for ARDS have supportive effects, making them inefficient. New methods such as stromal cell therapy are needed for this problem. METHODS: This research was performed with ten New Zealand rabbits in two groups. Bone marrow aspiration was performed on the treated group, and mesenchymal stem cells were isolated and cultured. The experimental model of ARDS was induced using LPS from Escherichia coli strain O55:B5. Then, 1010 bone marrow mesenchymal stem cells (BM-MSCs) were autologously transplanted intrapulmonary in the treatment group, and 1-2 ml of PBS in the control group. The clinical signs, computed tomographic (CT) scans, echocardiography, blood gas analysis, complete blood count, and cytokine levels were measured before and at 3, 6, 12, 24, 48, 72, and 168 h after BM-MSC transplant. Finally, the rabbits were killed, and histopathological examination was performed. RESULTS: The results showed that BM-MSCs decreased the severity of clinical symptoms, the number of white blood cells and heterophils in the blood, the total cell count, and number of heterophils and macrophages in bronchoalveolar lavage, and balanced the values of arterial blood gases (increase in partial pressure of oxygen and O2 saturation and decrease in the partial pressure of carbon dioxide). They also downregulated the tumor necrosis factor (TNF)-α and interleukin (IL)-6 concentrations and increased the IL-10 concentrations at different times compared with time 0 and in the control group, significantly. In the CT scan, a significant decrease in the Hounsfield units and total lung volume was found by echocardiography, and in comparing the two groups, a significant difference in the parameters was noticed. The histopathology demonstrated that the BM-MSCs were able to reduce the infiltration of inflammatory cells and pulmonary hemorrhage and edema. CONCLUSIONS: This study indicated that BM-MSCs play a significant role in the repair of lung injury.

Mechanical and Chemical Predifferentiation of Mesenchymal Stem Cells Into Cardiomyocytes and Their Effectiveness on Acute Myocardial Infarction.

Myocardial infarction is one of the leading causes of death all over the world. Mesenchymal stem cells (MSCs) transplantation has shown a promising potential to recovery of ischemic heart disease due to their capability in differentiating into cardiac cells. However, various investigations have been performed to optimize the efficacy of cardiac cell therapy in recent years. Here, we sought to interrogate the effect of autologous transplantation of undifferentiated and predifferentiated adipose and bone marrow-derived MSCs in a rabbit model of myocardial infarction and also to investigate whether cardiac function could be improved by mechanically induced MSCs via equiaxial cyclic strain. The two sources of MSCs were induced toward cardiomyocyte phenotype using mechanical loading and chemical factors and thereafter injected into the infarcted myocardium of 35 rabbits. Echocardiography and histopathology studies were used to evaluate cardiac function after 2 months. The results demonstrated significant scar size reduction and greater recovery of left ventricle ejection fraction after transplantation of predifferentiated cells, though the differences were not significant when comparing mechanically with chemically predifferentiated MSCs. Thus, although there was no significant improvement in infarcted myocardium between chemically and mechanically predifferentiated MSCs, mechanically induced cells are more preferred due to lack of any chemical intervention and cost reasonableness in their preparation method. Outcomes of this study may be useful for developing future therapeutic strategies, however long-term assessments are still required to further examine their effectiveness.

Generation of Lung and Airway Epithelial Cells from Embryonic Stem Cells In Vitro.

The epithelium covers the internal surface of the lung airways and is composed of numerous cell types that arise from the anterior foregut endoderm. The ability to generate pulmonary cells from embryonic stem cells will allow study of lung development, drug screening, regenerative medicine, and modeling of lung diseases. The aim of this review is to describe the methods used to produce lung cells in vitro, based on mechanisms of lung development in during embryology.

Preparation and Evaluation of Contact Lenses Embedded with Polycaprolactone-Based Nanoparticles for Ocular Drug Delivery.

To improve the efficiency of topical ocular drug administration, we focused on development of a nanoparticles loaded contact lens to deliver the hydrophobic drug over a prolonged period of time. The cross-linked nanoparticles based on PCL (poly ε-caprolactone), 2-hydroxy ethyl methacrylate (HEMA), and poly ethylene glycol diacrylate (PEG-DA) were prepared by surfactant-free miniemulsion polymerization. The lens material was prepared through photopolymerization of HEMA and N-vinylpyrrolidone (NVP) using PEG-DA as the cross-linker. Effects of nanoparticles loading on critical contact lens properties such as transparency, water content, modulus and ion and oxygen permeabilities were studied. Nanoparticles and hydrogel showed high viability, indicating the absence of cytotoxicity and stimulatory effect. Drug release studies revealed that the hydrogel embedded with nanoparticles released the drug for a period of 12 days. The results of this study provide evidence that nanoparticles loaded hydrogels could be used for extended delivery of loteprednol etabonate and perhaps other drugs.

Functional synergy of anti-mir221 and nanohydroxyapatite scaffold in bone tissue engineering of rat skull.

An appropriate cell source, effective cell modification, and proper supportive matrices are the main bases of tissue engineering. The effectiveness of anti-mir221 or hydroxyapatite (HA) in improving the osteogenic differentiation of mesenchymal stem cells (MSCs) has been reported previously. Herein, simultaneous application of these osteogenic inducers was investigated in vivo. The Poly-caprolactone (PCL)/HA nanofibers were characterized using contact angle measurement, tensile test, Fourier transform infrared spectroscopy, and electron microscopy. Rat MSCs were isolated, characterized and transfected with anti-mir221. The rats were divided into 4 groups and an 8 mm defect were created in the mid-calvaria of each rat by trephine bur. Group 1 received (PCL)/HA nanofibers, group 2 received (PCL)/HA nanofibers plus autologous MSCs, group 3 received (PCL)/HA nanofibers plus MSCs transfected with anti-mir221, and group 4 rats were left empty as an additional control group. Histomorphometric and radiomorphometric evaluation after 4 and 8 weeks revealed more new bone formation in the cell-treated groups compared to the scaffold alone group. There was evidence for a combination of increased osteoclasts and osteoblast vascular lake containing red blood cells in the anti-mir221 transfected group. New bone penetration into the scaffolds empirically demonstrated the capability of this combination for efficient osteointegration. Altogether, the co-application of HA and anti-mir221 transfected cells can enhance bone healing of the rat skull.

The effect of low-level laser therapy (810 nm) on root development of immature permanent teeth in dogs.

Traumatic injuries and dental caries can be a big challenge to immature teeth. In these cases, the main purpose of treatment is to maintain the pulp vitality. The purpose of this study was to investigate the effect of low-level laser therapy on accelerating the rate of dentinogenesis in pulpotomy of immature permanent teeth (apexogenesis). Three dogs, 4-6 months old, were used in this study. One jaw in each dog was randomly assigned to laser irradiation group. All selected teeth were pulpotomized with mineral trioxide aggregate (MTA) and restored with amalgam. In the laser group, the Ga-Al-As laser (810 nm, 0.3 W, 4 J/cm(2), 9 s) was used on buccal and lingual gingiva of each tooth in 48 h intervals for 2 weeks. In order to observe the newly formed dentine, tetracycline was injected on the 1st, 3rd, 7th, and 14th day after the operation. Then, ground sections of teeth were observed under a fluorescence microscope. The data was analyzed with Generalized Estimating Equations (GEE) test. The mean distance between the lines of tetracycline formed on the 1st and 14th day was significantly higher in the laser group (P = 0.005). Within the limitation of this study, irradiation of Ga-Al-As laser (810 nm) can accelerate the rate of dentinogenesis in apexogenesis of immature permanent teeth with MTA in dogs.

Comparing the effect of equiaxial cyclic mechanical stimulation on GATA4 expression in adipose-derived and bone marrow-derived mesenchymal stem cells.

Myocardium is prone to mechanical stimuli among which pulsatile blood flow exerts both radial and longitudinal strains on the heart. Recent studies have shown that mechanical stimulation can notably influence regeneration of cardiac muscle cells. GATA4 is a cardiac-specific transcription factor that plays an important role in late embryonic heart development. Our study aimed at investigating the effect of equiaxial cyclic strain on GATA4 expression in adipose-derived (ASCs) and bone marrow-derived (BMSCs) mesenchymal stem cells. For this reason, both ASCs and BMSCs were studied in four distinct groups of chemical, mechanical, mechano-chemical and negative control. According to this categorisation, the cells were exposed to cyclic mechanical loading and/or 5-azacytidine as the chemical factor. The level of GATA4 expression was then quantified using real-time PCR method on the first, fourth and seventh days. The results show that: (1) equiaxial cyclic stimulation of mesenchymal stem cells could promote GATA4 expression from the early days of induction and as it went on, its combination with chemical factor elevated expression; (2) cyclic strain could accelerate GATA4 expression compared to the chemical factor; (3) in this regard, these results indicate a higher capacity of ASCs than BMSCs to express GATA4.

Pre-vascularized porous gelatin-coated ß-tricalcium phosphate scaffolds for bone regeneration: an in vivo and in vitro investigation.

Vascularization is vital in bone tissue engineering, supporting development, remodeling, and regeneration. Lack of vascularity leads to cell death, necessitating vascularization strategies. Angiogenesis, forming new blood vessels, provides crucial nutrients and oxygen. Pre-vascularized gelatin-coated ß-tricalcium phosphate (G/ß-TCP) scaffolds show promise in bone regeneration and vascularization. Our study evaluates G/ß-TCP scaffolds' osteogenic and angiogenic potential in vitro and a canine model with vascular anastomosis. Channel-shaped G/ß-TCP scaffolds were fabricated using foam casting and sintering of a calcium phosphate/silica slurry-coated polyurethane foam, then coated with cross-linked gelatin. Buccal fat pad-derived stem cells (BFPdSCs) were seeded onto scaffolds and assessed over time for adhesion, proliferation, and osteogenic capacity using scanning electron microscopy (SEM), 4,6-diamidino-2-phenylindole (DAPI) staining, Alamar blue, and alkaline phosphatase (ALP) assays. Scaffolds were implanted in a canine model to evaluate osteogenesis and angiogenesis by histology and CT scans at 12 wk. Our studies showed preliminary results for G/ß-TCP scaffolds supporting angiogenesis and bone regeneration. In vitro analyses demonstrated excellent proliferation/viability, with BFPdSCs adhering and increasing on the scaffolds. ALP activity and protein levels increased, indicating osteogenic differentiation. Examination of tissue samples revealed granulation tissue with a well-developed vascular network, indicating successful angiogenesis and osteogenesis was further confirmed by a CT scan. In vivo, histology revealed scaffold resorption. However, scaffold placement beneath muscle tissue-restricted bone regeneration. Further optimization is needed for bone regeneration applications.

Evaluating osteogenic potential of a 3D-printed bioceramic-based scaffold for critical-sized defect treatment: an in vivo and in vitro investigation.

The integration of precision medicine principles into bone tissue engineering has ignited a wave of research focused on customizing intricate scaffolds through advanced 3D printing techniques. Bioceramics, known for their exceptional biocompatibility and osteoconductivity, have emerged as a promising material in this field. This article aims to evaluate the regenerative capabilities of a composite scaffold composed of 3D-printed gelatin combined with hydroxyapatite/tricalcium phosphate bioceramics (G/HA/TCP), incorporating human dental pulp-derived stem cells (hDPSCs). Using 3D powder printing, we created cross-shaped biphasic calcium phosphate scaffolds with a gelatin layer. The bone-regenerating potential of these scaffolds, along with hDPSCs, was assessed through in vitro analyses and in vivo studies with 60 rats and critical-sized calvarial defects. The assessment included analyzing cellular proliferation, differentiation, and alkaline phosphatase activity (ALP), and concluded with a detailed histological evaluation of bone regeneration. Our study revealed a highly favorable scenario, displaying not only desirable cellular attachment and proliferation on the scaffolds but also a notable enhancement in the ALP activity of hDPSCs, underscoring their pivotal role in bone regeneration. However, the histological examination of calvarial defects at the 12-wk mark yielded a rather modest level of bone regeneration across all experimental groups. The test and cell group exhibited significant bone formation compared to all other groups except the control and cell group. This underscores the complexity of the regenerative process and paves the way for further in-depth investigations aimed at improving the potential of the composite scaffolds.

Bioinspired wound dressing: Investigating coelomic fluid-enhanced chitosan/polyvinyl alcohol nanofibers.

The electrospun mats consisting of integrated coelomic fluid (CF) and chitosan (Chs) into polyvinyl alcohol (PVA) nanofibers were produced and evaluated for use as wound dressings. CF was obtained from earthworms (Eisenia andrei (Fetida)) using an electric shock method, while Chs was chemically produced from shrimp chitin and then characterized using titration, Fourier transform infrared (FT-IR) spectroscopy, and viscometry. The wound dressings with different CF contents were evaluated for their antibacterial, antioxidant, and cell viability properties. The dressings infused with CF showed significantly higher antibacterial and antioxidant activity, as well as improved cell viability compared to the control without CF. In vivo studies using adult Wistar albino rats showed that the Chs/PVA/CF wound dressings promoted wound healing and re-epithelialization. Moreover, histological examinations of the injuries coated with Chs/PVA/CF displayed improved re-epithelialization. These results suggest that the Chs/PVA/CF nanofiber has the potential for use as a wound dressing material.

Effect of Intra-ovarian Injection of Mesenchymal Stem Cells or its Conditioned Media on Repeated OPU-IVEP Outcomes in Jersey Heifers and Its Relationship with Follicular Fluid Inflammatory Markers.

Background: Repeated Ovum Pick Up (OPU) could have a detrimental effect on ovarian function, reducing In Vitro Embryo Production (IVEP). The present study examined the therapeutic effect of adipose-derived Mesenchymal Stem Cells (MSCs) or its Conditioned Medium (ConM) on ovarian trauma following repeated OPU. Resolvin E1 (RvE1) and Interleukin-12 (IL-12) were investigated as biomarkers. Methods: Jersey heifers (n=8) experienced 11 OPU sessions including 5 pre-treatment and 6 treatment sessions. Heifers received intra-ovarian administration of MSCs or ConM (right ovary) and Dulbecco's Modified Phosphate Buffer Saline (DMPBS; left ovary) after OPU in sessions 5 and 8 and 2 weeks after session 11. The concentrations of RvE1 and IL-12 in follicular fluid was evaluated on sessions 1, 5, 6, 9, and 4 weeks after session 11. Following each OPU session, the IVEP parameters were recorded. Results: Intra-ovarian administration of MSCs, ConM, and DMPBS did not affect IVEP parameters (p>0.05). The concentration of IL-12 in follicular fluid increased at the last session of pre-treatment (Session 5; p<0.05) and remained elevated throughout the treatment period. There was no correlation between IL-12 and IVEP parameters (p>0.05). However, RvE1 remained relatively high during the pre-treatment and decreased toward the end of treatment period (p<0.05). This in turn was associated with decline in some IVEP parameters (p<0.05). Conclusion: Intra-ovarian administration of MSCs or ConM during repeated OPU did not enhance IVEP outcomes in Bos taurus heifers. The positive association between RvE1 and some of IVEP parameters could nominate RvE1 as a promising biomarker to predict IVEP parameters following repeated OPU.

Biological evaluation of polyvinyl alcohol hydrogel crosslinked by polyurethane chain for cartilage tissue engineering in rabbit model.

Polyvinyl alcohol (PVA) hydrogel chains were crosslinked by urethane pre-polymer (PPU) in order to fabricate a new substitute for cartilage lesions. The microscopy images showed that the cultured chondrocytes had spherical morphology on PVA-PPU sample after 4 weeks of isolation in vitro. The alcian blue and safranin O staining proved the presence of proteoglycan on the surface of PVA-PPU sample secreted by cultured chondrocytes. This was confirmed by the detection of sulfate ions in the wavelength dispersive X-ray (WDX) analysis. In addition, the expression of collagen type II and aggrecan were observed in chondrocytes cultured on PVA-PPU by RT-PCR. Moreover, the implantation of the PVA-PPU sample with autologous cultured chondrocytes revealed the formation of neocartilage tissue in a rabbit model during 12 weeks follow up. In conclusion, the results verified that isolated chondrocytes cultured on PVA-PPU retain their original phenotype and this composition can be considered as promising substrate for cartilage tissue engineering.

Healing of extraction sockets and augmented alveolar defects following 1-year treatment with bisphosphonate.

To assess the effect of bisphosphonates on healing of extraction sockets and augmented alveolar defects, 12 adult female mongrel dogs were assigned to 2 experimental groups and a control group. The experimental groups received oral alendronate (ALN, 3.5 mg/kg/wk) or IV pamidronate (PAM, 1 mg/kg/wk) for 12 months. Animals were randomly tested for serum C-terminal telopeptide of collagen I (CTx). The right first and second premolars were extracted. After 8 weeks, extraction sites were evaluated for healing. Subsequently, 3-wall defects were created in ridges and filled with human mineralized cortical particulate bone. Two months post-augmentation, animals were sacrificed and mandibles were collected for cone-beam computed tomography (CBCT) and histomorphometric appraisal. The obtained data were compared using 1-way ANOVA test. CTx test results in both experimental groups were comparable (<10 pg/mL) but lower than that of the control group (minimum 159.2 pg/mL). Two months post-extraction, bone sequestra were noticed in extraction sites in BP-treated groups, involving the entire alveolar bone in the PAM group and the upper rim of the alveoli in the ALN group. Histologically, bone sequestra from the PAM group demonstrated empty osteocyte lacunae, while in the ALN group areas of necrotic bone along with evidence of active bone remodeling was distinguished. Eight weeks post-augmentation, the experimental groups showed no evidence of bone formation in the augmented area, while bone formation ratio was measured to be 18.32% in the control group. The mean amount of pixel intensity calculated from the CBCT images of the ALN, PAM, and control group was 113.69 ± 11.04, 124.94 ± 4.72, and 113.69 ± 6.63, respectively. Pixel intensity in PAM-treated group was significantly higher than both other groups. This study demonstrated that 1-year treatment with ALN/PAM was associated with impairment of post-extraction and post-augmentation bone healing in dogs.

Kaempferol-loaded bioactive glass-based scaffold for bone tissue engineering: in vitro and in vivo evaluation.

Due to the increasing prevalence of bone disorders among people especially in average age, the future of treatments for osseous abnormalities has been illuminated by scaffold-based bone tissue engineering. In this study, in vitro and in vivo properties of 58S bioactive glass-based scaffolds for bone tissue engineering (bare (B.SC), Zein-coated (C.SC), and Zein-coated containing Kaempferol (KC.SC)) were evaluated. This is a follow-up study on our previously published paper, where we synthesized 58S bioactive glass-based scaffolds coated with Kaempferol-loaded Zein biopolymer, and characterized from mostly engineering points of view to find the optimum composition. For this aim, in vitro assessments were done to evaluate the osteogenic capacity and biological features of the scaffolds. In the in vivo section, all types of scaffolds with/without bone marrow-derived stem cells (BMSC) were implanted into rat calvaria bone defects, and potential of bone healing was assessed using imaging, staining, and histomorphometric analyses. It was shown that, Zein-coating covered surface cracks leading to better mechanical properties without negative effect on bioactivity and cell attachment. Also, BMSC differentiation proved that the presence of Kaempferol caused higher calcium deposition, increased alkaline phosphatase activity, bone-specific gene upregulation in vitro. Further, in vivo study confirmed positive effect of BMSC-loaded KC.SC on significant new bone formation resulting in complete bone regeneration. Combining physical properties of coated scaffolds with the osteogenic effect of Kaempferol and BMSCs could represent a new strategy for bone regeneration and provide a more effective approach to repairing critical-sized bone defects.

In vitro investigation of canine periodontal ligament-derived mesenchymal stem cells: A possibility of promising tool for periodontal regeneration.

Objectives: Recent investigations indicate that canine periodontal ligament-derived stem cells (cPDLSCs) may reveal a reliable strategy for repair of periodontal tissues via cell-based tissue engineering approaches. Due to limited research, this study aimed to demonstrate the phenotypic characterization of cPDLSc in comparison with canine bone marrow-derived mesenchymal stem cells (cBMSCs) in vitro. Methods: Mesenchymal stem cells (MSCs) were obtained from PDL and BM of five male adult Mongrel dogs. In vitro isolation and expansion as well as biologic characterization including colony unit formation (CFU), osteogenic and adipogenic differentiation, flow cytometric analysis of CD34 and CD44, and RT-PCR of alkaline phosphatase (ALP), osteocalcin (OCN), periostin (POSTN) and S100A4 were performed. Furthermore, electron microscopy analysis was done to complement the comparative research. Results: CFU assay revealed that colonies of cPDLSCs presented 70% confluency with a more finite lifespan than BM-MSCs, showing a significant increase in cPDLSCs. Both types of MSCs showed osteogenic and adipogenic phenotypic characterized with clusters of mineralized depositions and lipid vacuoles, respectively. Both types of MSCs expressed CD44 with limited expression of CD34. RT-PCR of cPDLSCs revealed that expression of ALP, POSTN, OCN and S100A4 genes were significantly higher than those of BMSCs. In addition, comparison of SEM and revealed that cPDLSCs expressed more extracellular collagen fibers. Conclusions: The current study indicated that cPDLSCs show potency as a novel cellular therapy for periodontal regeneration a large animal model.

Stereological study of testes following experimentally-induced unilateral cryptorchidism in rats.

OBJECTIVE: Cryptorchidism is one of the main causes of infertility and can result in testicular cancer. This study aimed to present quantitative data on the damage caused by cryptorchidism using stereological analysis. METHODS: Thirty newborn rats were randomly divided into control and experimental groups. The experimental group underwent surgery to induce unilateral cryptorchidism in the left testis, whereas the control group underwent a sham surgical procedure 18 days after birth. The testes were removed at designated time points (40, 63, and 90 days after birth) for stereological evaluation and sperm analysis. Total testicular volume, interstitial tissue volume, seminiferous tubule volume and length, and seminiferous epithelium volume and surface area were measured. Other parameters, such as sperm count, sperm morphology, and sperm tail length, were also examined. RESULTS: Statistically significant differences (p<0.05) were observed between the experimental and the control groups at different ages regarding the volumes of various parameters, including the surface area of the germinal layer, the length of the seminiferous tubules, sperm count, and sperm morphology. However, no significant differences were observed in the epithelial volume and the sperm tail length of the groups. CONCLUSION: Given the substantial effect of cryptorchidism on different testicular parameters, as well as the irreversible damage it causes in the testes, it is important to take this abnormality seriously to prevent these consequences.