Amino acid surface modified bioglass: A candidate biomaterial for bone tissue engineering1.

Bioglasses are solid materials consisted of sodium oxide, calcium oxide, silicon dioxide and phosphorus in various proportions and have used in bone tissue engineering. There have been ongoing efforts to improve the surface properties of bioglasses to increase biocompatibility and performance. The aim of the present study is to modify the bioglass surface with an amino acid mixture consisting of arginine, aspartic acid, phenylalanine, cysteine, histidine and lysine, to characterize the surface, and to evaluate the performance and biocompatibility in vitro and in vivo. The untreated bioglass, bioglass kept in simulated body fluid (SBF), and modified bioglass were used in further evaluation. After confirmation of the surface modification with FT-IR analyses and SEM analyses, MC3T3-E1 preosteoblasts adhesion on the surface was also revealed by SEM. The modified bioglass had significantly higher ALP activity in colorimetric measurement, rate of calcium accumulations in Alizarin red s staining, lower rate of cell death in Annexin-V/PI staining to determine apoptosis and necrosis. Having higher cell viability rate in MTT test and absence of genotoxicity in micronucleus test (OECD 487), the modified bioglass was further confirmed for biocompatibility in vitro. The results of the rat tibial defect model revealed that the all bioglass treatments had a significantly better bone healing score compared to the untreated negative control. However, the modified bioglass exhibited significantly better bone healing efforts especially during the first and the second months compared to the other bioglass treatment treatments. As a result, the amino acid surface modification of bioglasses improves the surface biocompatibility and osteogenic performance that makes the amino acid modified bioglass a better candidate for bone tissue engineering. RESEARCH HIGHLIGHTS: Bioglass surface modification with amino acids contributes to bioglass-tissue interaction with an improved cell attachment. Modified bioglass increases in vitro Alp activity and calcium accumulation, and also positively affects cell behavior by supporting cell adaptation. Bioglass exerts osteogenic potential in vivo especially during early bone healing.

The impact of bone marrow-derived mesenchymal stem cells on experimental testiculartorsion in rats.

BACKGROUND: The aim of this study was to investigate the healing effects of bone marrow-derived mesenchymal stem cells (BMMSCs) on experimental testicular torsion in rats. METHODS: Three groups consisting of 10 Wistar albino rats were created. In Group I, the left testicle was explored and relocated in the scrotum without any attempt to modify it. In Group II, the left testicle underwent torsion for three h and then was detorsed and relocated. In Group III, in addition to torsion and detorsion, BM-MSCs were administered intratesticularly. The rats were sacrificed on the seventh day, and the healing status of the testicles was investigated with histopathological and biochemical analyses. BM-MSC involvement was investigated by immunofluorescence microscopy. Statistical analysis was performed using SPSS 15.0. A p-value < 0.05 was considered statistically significant for all variables. RESULTS: Immunofluorescence microscopy showed that BM-MSCs were located around the Leydig cells in Group III. Under light microscopy, the mean Johnsen Score of Group III was significantly higher than that of Group II (p = 0.035). The interleukin-10 (IL-10) level was significantly higher in Group III compared to Group II (p = 0.003). While the malondialdehyde (MDA) values in Group I (the control group) were lower than in the other groups (p = 0.037), the superoxide dismutase (SOD) values were similar (p = 0.158). Although there was no statistically significant difference between Group II and Group III in terms of MDA, it was lower in Group III. Although the tissue SOD levels were higher in Group III than in Group II, the difference was not statistically significant. DISCUSSION: : This study has demonstrated that BM-MSCs significantly corrected the Johnsen Score and increased anti-inflammatory cytokine levels after testicular torsion. BM-MSCs can be used in testicular torsion as supportive therapy to minimize tissue damage.

The impact of bone marrow-derived mesenchymal stem cells on experimental testicular torsion in rats.

Background/aim: The aim of this study was to investigate the healing effects of bone marrow-derived mesenchymal stem cells (BM-MSCs) on experimental testicular torsion in rats. Materials and methods: Three groups consisting of 10 Wistar albino rats were created. In Group I, the left testicle was explored and relocated in the scrotum without any attempt to modify it. In Group II, the left testicle underwent torsion for three h and then was detorsed and relocated. In Group III, in addition to torsion and detorsion, BM-MSCs were administered intratesticularly. The rats were sacrificed on the seventh day, and the healing status of the testicles was investigated with histopathological and biochemical analyses. BM-MSC involvement was investigated by immunofluorescence microscopy. Statistical analysis was performed using SPSS 15.0. A p-value < 0.05 was considered statistically significant for all variables. Results: Immunofluorescence microscopy showed that BM-MSCs were located around the Leydig cells in Group III. Under light microscopy, the mean Johnsen Score of Group III was significantly higher than that of Group II (p = 0.035). The interleukin-10 (IL-10) level was significantly higher in Group III compared to Group II (p = 0.003). While the malondialdehyde (MDA) values in Group I (the control group) were lower than in the other groups (p = 0.037), the superoxide dismutase (SOD) values were similar (p = 0.158). Although there was no statistically significant difference between Group II and Group III in terms of MDA, it was lower in Group III. Although the tissue SOD levels were higher in Group III than in Group II, the difference was not statistically significant. Conclusion: This study has demonstrated that BM-MSCs significantly corrected the Johnsen Score and increased anti-inflammatory cytokine levels after testicular torsion. BM-MSCs can be used in testicular torsion as supportive therapy to minimize tissue damage.

Contribution of Bone Marrow-Derived Mesenchymal Stem Cells to Healing of Pulmonary Contusion-Created Rats.

BACKGROUND: The most common thoracic injury in children, resulting in trauma, is pulmonary contusion (PC). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are used in wound healing and many other diseases. This study aims to examine the effects of BM-MSCs on PC healing in rats. MATERIALS AND METHODS: A total of 45 male Wistar albino rats were used. Four groups were formed. BM-MSCs were labeled with the green fluorescent protein. PC was observed in the control group. In group II, PC occured and left to spontaneous healing. In group III, PC formed and BM-MSCs were given. In group IV, BM-MSCs were given without PC formation. Subjects were sacrificed 1 week later. Whether there was any difference in terms of BM-MSC involvement and lung injury score was investigated. Statistical analysis was performed using the Statistical Package for Social Sciences (SPSS), version 17.0, software (SPSS Inc., Chicago, IL), and p value of <0.05 was considered statistically significant. RESULTS: BM-MSCs were collected much more in the lungs in group III than in group IV. Group III had a lower lung injury score value than group II. CONCLUSION: The greater involvement of the BM-MSCs in the injury site, and further reductions in lung injury score suggest that BM-MSCs are contributing to the healing of the injury. The use of BM-MSCs in risky patients with diffuse PC may be an alternative treatment to conventional methods.

Autologous stem cell-derived chondrocyte implantation with bio-targeted microspheres for the treatment of osteochondral defects.

BACKGROUND: Chondral injury is a common problem around the world. Currently, there are several treatment strategies for these types of injuries. The possible complications and problems associated with conventional techniques lead us to investigate a minimally invasive and biotechnological alternative treatment. Combining tissue-engineering and microencapsulation technologies provide new direction for the development of biotechnological solutions. The aim of this study is to develop a minimal invasive tissue-engineering approach, using bio-targeted microspheres including autologous cells, for the treatment of the cartilage lesions. METHOD: In this study, a total of 28 sheeps of Akkaraman breed were randomly assigned to one of the following groups: control (group 1), microfracture (group 2), scaffold (group 3), and microsphere (group 4). Microspheres and scaffold group animals underwent adipose tissue collection prior to the treatment surgery. Mesenchymal cells collected from adipose tissue were differentiated into chondrocytes and encapsulated with scaffolds and microspheres. Osteochondral damage was conducted in the right knee joint of the sheep to create an animal model and all animals treated according to study groups. RESULTS: Both macroscopic and radiologic examination showed that groups 3 and 4 have resulted better compared to the control and microfracture groups. Moreover, histologic assessments indicate hyaline-like cartilage formations in groups 3 and 4. CONCLUSION: In conclusion, we believe that the bio-targeted microspheres can be a more effective, easier, and safer approach for cartilage tissue engineering compared to previous alternatives.

Effects of Local Low-Dose Alendronate Injections Into the Distraction Gap on New Bone Formation and Distraction Rate on Distraction Osteogenesis.

Bisphosphonates that constrain bone resorption have a direct effect on osteoclast function. In this experimental study, the effects of low-dose local alendronate injections on the distraction gap (DG) in rabbit mandible at 2 different rates were evaluated.The experimental study was conducted on 20 male, New Zealand white rabbits. The animals were divided into 3 experimental groups and 1 control group. Group 1 consisted of animals with distraction at the rate of 1 mm/day, receiving postoperative local low-dose alendronate local injections into the DG. Group 2 consisted of animals with distraction at the rate of 2 mm/day, receiving postoperative 0.75 µg/kg of alendronate local injections into the DG. Group 3 consisted of animals with distraction at the rate of 2 mm/day, receiving postoperative 0.2 mL local saline injections into the DG. Group 4 consisted of animals with distraction at the rate of 1 mm/day, receiving postoperative 0.2 mL local saline injections into the DG. All the injections were performed immediately postoperatively and for all groups at 1, 2, 3, and 4 weeks following surgery. The distraction zones were evaluated using dual-energy X-ray absorptiometry and histological analysis.Histologically, bone healing was found to be significantly accelerated in Groups 1 and 4 compared with Groups 2 and 3 (P < 0.05). Bone healing was superior in Group 1 and the difference was statistically significant compared with Group 4. There was a significant increase in mean bone mineral density in the 1 mm daily rate groups (Groups 1 and 4) compared with the 2 mm daily rate groups (Groups 2 and 3) (P < 0.05).Local low-dose alendronate injections could be an effective way for improving bone formation in distraction osteogenesis. Furthermore, the results of this study did not support the hypothesis that injections of local low-dose alendronate may allow 2 mm/day instead of 1 mm/day of elongation in the rabbit mandible.

In-vivo efficacy of toltrazuril on experimentally induced Toxoplasma gondii tissue cysts in lambs: a novel strategy for prevention of human exposure to meat-borne toxoplasmosis.

The aim of the present study was to investigate in vivo efficacy of toltrazuril on Toxoplasma gondii tissue cysts following induction of chronic toxoplasmosis in 4-week-old lambs (n=27) by inoculation of 1×10(5) T. gondii ME 49 strain oocysts (day 0). Beginning at the 15th day after inoculation, lambs in Group T20 and Group T40 were given toltrazuril orally 2 times, once every week (Baycox 5%, Bayer Animal Health) at a dose of 20 mg/kg and 40 mg/kg, respectively. Positive control (PC) lambs were not given any therapy, and 2 clinically healthy non-infected lambs were used as negative controls (Group NC). Two out of 9 lambs in PC group (oocyst inoculated but non-treated) were killed on toltrazuril treatment days (day 15 and 22) to evaluate the tissue cyst presence in their muscles. On day 90, the remaining 25 lambs were necropsied, and samples from the brain and 11 different muscle groups were collected. The tissues were examined for the presence of tissue cysts by histopathology, immunohistochemistry, nested-PCR and percoll gradient centrifugation. Anti-T. gondii antibodies were screened by IFAT throughout the experiment. The increased T. gondii seropositivity beginning from the 15th day of inoculation remained steady at Day 45 and Day 90 in Groups PC while it was significantly lower at Day 90 in toltrazuril receiving groups. In toltrazuril treated groups, histopathological findings included degenerative changes in the cyst wall, complete macrophage invasion to the cysts, and reduction or removal of the cysts in toto. Four out of 9 lambs (44.4%) in both toltrazuril treated group (Group T20 and T40) did not contain tissue cyst in any examined tissues while all positive control animals had T. gondii tissue cysts at least in one muscle group. The toltrazuril treatment efficacy on the cyst presence was determined as 44.4%. The number of the cysts in the musculature was significantly different between non-treated and toltrazuril treated lambs (X(2)=6.613; p=0.037). For the total number of cysts, the positive control lambs had higher number of cysts compared to both toltrazuril treated lambs (T20 and T40) (X(2)=5.629; p=0.018 and X(2)=5.629; p=0.018, respectively) while there were no differences between Group T20 and Group T40 (X(2)=0.000; p=1.000). According to PCR results, the brain and M. semitendinosus were positive in all 7 control lambs while 12 out of 18 lambs were positive in toltrazuril treated lambs. In conclusion, the results are promising as the toltrazuril treated lambs had markedly less parasite counts compared to those of untreated lambs. Further research should be conducted to reveal if toltrazuril treatment in sheep could be used as a strategy to minimize the cyst exposure of humans through consumption of raw or undercooked mutton.

Cerebellum progesterone concentration decreased in canine distemper virus infection.

Progesterone has neuroprotective effects including augmentation of myelination in the central and peripheral nervous system. This study was designed to determine if demyelinating lesions in the cerebellum resulting from canine distemper virus (CDV) infection are associated with progesterone levels. Progesterone was measured using radioimmunoassay in samples of the cerebellum, corpus callosum, medulla oblongata, parietal, frontal, temporal, and occipital cortices as well as cerebrospinal fluid (CSF) and plasma collected from ten CDV infected and six non-infected dogs. The cerebellum progesterone level was significantly different between CDV infected (0.66+/-0.09 ng/g) and control dogs (1.14+/-0.09 ng/g) (p<0.001); however, no difference was observed for the other CNS regions, plasma and CSF (p>0.05). The cerebellum progesterone level was also significantly different between acute (0.71+/-0.0 5 ng/g) and chronic cases (0.61+/-0.09 ng/g) (p<0.05). The CDV infected cerebella were also categorized histopathologically according to the severity of demyelinating lesions as mild (n=5), moderate (n=2), or severe (n=3) among which the cerebellum progesterone level was significantly different (p<0.05). Progesterone concentration was 0.71+/-0.05 ng/g in mild, 0.65+/-0.10 ng/g in moderate, and 0.56+/-0.07 ng/g in severe cases. In conclusion, progesterone concentration decreases in the cerebellum in CDV infection and the severity of demyelinating lesions is the greatest in cerebella with the lowest progesterone concentrations. The results suggest that local impairment of progesterone metabolism may be associated with the initiation and progression of cerebellar lesions in CDV infection.

Immunolocalization of 3beta-hydroxysteroid dehydrogenase in normal and hyperplastic ram prostates.

The enzyme 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is essential in the synthesis of all steroids by cleaving dehydroepiandrosterone to androstenedione. In the present study, 3beta-HSD immunoreactivity was investigated in the prostate of Akkaraman breed rams aged older than 3 years. Five normal and five hyperplastic ram prostates were processed for immunohistochemistry. Prostate hyperplasia was determined by histopathological evaluation of 375 ram prostate and confirmed with significantly (P<0.01) increased number of cells expressing proliferating cell nuclear antigen (PCNA) immunoreactivity in the glandular epithelia. The 3beta-HSD immunoreactivity with a variable intensity and pattern of distribution was present in the glandular epithelia and endothelia of blood vessels in normal and hyperplastic ram prostates. While immunoreactivity was focally present in some glands, some sections had a homogenous distribution. The presence of 3beta-HSD immunoreactivity indicates that steroids are locally synthesized in the ram prostate. No differences in the distribution pattern of 3beta-HSD immunoreactivity and the percentage of immunoreactive cells were observed between normal and hyperplastic prostates (P>0.05), suggesting that locally produced steroids have little or no effect on the pathogenesis of the ram prostate hyperplasia which affects a very small proportion of the ram population (5 out of 375).