Effect of cilostazol on healing of achilles tendon ruptures: an experimental study on rats.

PURPOSE: This study aimed to evaluate whether cilostazol (phosphodiesterase III inhibitor) could enhance the healing of Achilles tendon ruptures in rats. MATERIALS AND METHODS: The Achilles tendons of 24 healthy male adult rats were incised and repaired. The rats were randomly allocated to cilostazol and control groups. The cilostazol group received daily intragastric administration of 50 mg/kg cilostazol for 28 days, while the control group did not receive any medication. The rats were sacrificed on the 30th day, and the Achilles tendon was evaluated for biomechanical properties, histopathological characteristics, and immunohistochemical analysis. RESULTS: All rats completed the experiment. The Movin sum score of the control group was significantly higher (p = 0.008) than that of the cilostazol group, with means of 11 ± 0.63 and 7.50 ± 1.15, respectively. Similarly, the mean Bonar score was significantly higher (p = 0.026) in the control group compared to the cilostazol group (8.33 ± 1.50 vs. 5.5 ± 0.54, respectively). Moreover, the Type I/Type III Collagen ratio was notably higher (p = 0.016) in the cilostazol group (52.2 ± 8.4) than in the control group (34.6 ± 10.2). The load to failure was substantially higher in the cilostazol group than in the control group (p = 0.034), suggesting that the tendons in the cilostazol group were stronger and exhibited greater resistance to failure. CONCLUSIONS: The results of this study suggest that cilostazol treatment significantly improves the biomechanical and histopathological parameters of the healing Achilles tendon in rats. Cilostazol might be a valuable supplementary therapy in treating Achilles tendon ruptures in humans. Additional clinical studies are, however, required to verify these outcomes.

Lentiviral Micro-dystrophin Gene Treatment into Late-stage mdx Mice for Duchenne Muscular Dystrophy Disease.

AIM: Duchenne Muscular Dystrophy (DMD) results in a deficiency of dystrophin expression in patient muscle fibers, leading to progressive muscle degeneration. Treatment of DMD has undertaken current transformation with the advancement of novel gene therapy and molecular biology techniques, which are secure, well-tolerated, and effective therapeutic approaches. INTRODUCTION: DMD gene therapies have mainly focused on young DMD patients as in vivo animal model trials have been performed in 0-1-month DMD mice. However, it has not yet been answered how micro-dystrophin encoding lentiviral treatment affects Dystrophin expression and DMD symptoms in 10-month mdx mice. METHODS: We planned to integrate the micro-Dystrophin gene sequence into the muscle cells by viral transfer, using micro-Dystrophin-encoding lentivirus to reduce the dystrophic pathology in late-stage dmd mice. The histopathological and physiological-functional regeneration activities of the lentiviralmicro- Dystrophin gene therapy methods were compared, along with changes in temporal Dystrophin expression and their functionality, toxicity, and gene expression level. RESULTS: Here, we showed that the micro-dystrophin transgene transfers intramuscularly and intraperitoneally in late-stage dmd-mdx-4cv mice restored dystrophin expression in the skeletal and cardiac muscle (p <0.001). Furthermore, motor performance analysis, including hanging and tracking tests, improved statistically significantly after the treatment (p <0.05). CONCLUSION: Consequently, this study suggests that patients in the late stages of muscular dystrophy can benefit from lentiviral micro-dystrophin gene therapies to present an improvement in dystrophic muscle pathology.

Effects of stem cells and amniotic fluid on uterus and ovaries in a rat model of abdominal adhesions: a controlled study

Objective: This study aimed to compare the effects of human umbilical cord mesenchymal stem cells (hUCMSCs), amniotic fluid (AF), and a combination of both on the uterus and ovaries in a rat model of abdominal adhesions. Material and Methods: This study was designed as a controlled study. Four groups, each consisting of six rats, were randomly formed. One group was designated as the control (CNT). hUCMSCs - applied (hUCSC), AF-applied (AMN), and a combination of both (hUCSC + AMN) were the experimental groups. All rats were given intraperitoneal talc powder to create adhesions. After 21 days, animals in experimental groups were further treated with hUCMSC, AF or a combination of these. Results: There was a statistically significant difference in primordial follicle count, endometrial gland number, and endometrial blood vessel count (p<0.05). AMN provided the best results in the endometrial vessel and primordial follicle count. The average endometrial gland count in AMN and hUCSC + AMN was similarly higher than CNT and hUCSC alone. Conclusion: There were significantly higher for counts for endometrial glands, endometrial blood vessels, and primordial follicles in the hUCSC, AMN and hUCSC + AMN groups compared to controls. Animals in the AMN group had the best result for endometrial vessel and highest primordial follicle count.

Gamma-irradiated SARS-CoV-2 vaccine candidate, OZG-38.61.3, confers protection from SARS-CoV-2 challenge in human ACEII-transgenic mice.

The SARS-CoV-2 virus caused the most severe pandemic around the world, and vaccine development for urgent use became a crucial issue. Inactivated virus formulated vaccines such as Hepatitis A and smallpox proved to be reliable approaches for immunization for prolonged periods. In this study, a gamma-irradiated inactivated virus vaccine does not require an extra purification process, unlike the chemically inactivated vaccines. Hence, the novelty of our vaccine candidate (OZG-38.61.3) is that it is a non-adjuvant added, gamma-irradiated, and intradermally applied inactive viral vaccine. Efficiency and safety dose (either 1013 or 1014 viral RNA copy per dose) of OZG-38.61.3 was initially determined in BALB/c mice. This was followed by testing the immunogenicity and protective efficacy of the vaccine. Human ACE2-encoding transgenic mice were immunized and then infected with the SARS-CoV-2 virus for the challenge test. This study shows that vaccinated mice have lowered SARS-CoV-2 viral RNA copy numbers both in oropharyngeal specimens and in the histological analysis of the lung tissues along with humoral and cellular immune responses, including the neutralizing antibodies similar to those shown in BALB/c mice without substantial toxicity. Subsequently, plans are being made for the commencement of Phase 1 clinical trial of the OZG-38.61.3 vaccine for the COVID-19 pandemic.

Is denervation surgery possible in the treatment of hallux rigidus? An anatomic study of cadaveric specimens.

OBJECTIVE: The aim of this study was to provide anatomic considerations in the first metatarsophalangeal joint (FMPJ) innervation and to evaluate the feasibility of the denervation surgery in the treatment of hallux rigidus. METHODS: In this cadaveric study, 14 fresh frozen cadaveric transtibial amputation specimens was used. For nerve dissection, dorsal and plantar longitudinal incision centered over the FMPJ were performed. Deep peroneal and dorsomedial cutaneous nerves were dissected in the dorsal aspect of the joint. Medial plantar nerve branches, medial and lateral hallucal nerves, were dissected in the plantar aspect of the joint. The presence, number, and location of articular branches to the FMPJ capsule were recorded. Dorsal and plantar incision length for proper dissection were also recorded. RESULTS: Nerve dissection of the 14 specimens revealed the following number of articular branches from the relevant nerves: 14 from dorsomedial cutaneous nerves, 11 from deep peroneal nerves, 6 from medial hallucal nerve, and 5 from lateral hallucal nerve. Dorsal incision mean length was 60.53 (range, 42.48-85.12) mm, and the plantar incision mean length was 88.08 (range, 77.32-111.21) mm. CONCLUSION: Evidence from this study has shown that partial dorsal denervation of the FMPJ may be a technically feasible procedure along with the presence of superficially easily dissected nerves with relatively small incision. LEVEL OF EVIDENCE: Level 5.

Preclinical efficacy and safety analysis of gamma-irradiated inactivated SARS-CoV-2 vaccine candidates.

COVID-19 outbreak caused by SARS-CoV-2 created an unprecedented health crisis since there is no vaccine for this novel virus. Therefore, SARS-CoV-2 vaccines have become crucial for reducing morbidity and mortality. In this study, in vitro and in vivo safety and efficacy analyzes of lyophilized vaccine candidates inactivated by gamma-irradiation were performed. The candidate vaccines in this study were OZG-3861 version 1 (V1), an inactivated SARS-CoV-2 virus vaccine, and SK-01 version 1 (V1), a GM-CSF adjuvant added vaccine. The candidate vaccines were applied intradermally to BALB/c mice to assess toxicity and immunogenicity. Preliminary results in vaccinated mice are reported in this study. Especially, the vaccine models containing GM-CSF caused significant antibody production with neutralization capacity in absence of the antibody-dependent enhancement feature, when considered in terms of T and B cell responses. Another important finding was that the presence of adjuvant was more important in T cell in comparison with B cell response. Vaccinated mice showed T cell response upon restimulation with whole inactivated SARS-CoV-2 or peptide pool. This study shows that the vaccines are effective and leads us to start the challenge test to investigate the gamma-irradiated inactivated vaccine candidates for infective SARS-CoV-2 virus in humanized ACE2 + mice.

Is melatonin, leptin or their combination more effective on oxidative stress and folliculogenesis in the obese rats?

In this study, we evaluated the effects of melatonin (Mel), leptin (Lep) or melatonin and leptin treatment on ovaries in control and obese rats. The animals were divided into control (NC), melatonin (NM), leptin (NL), melatonin-leptin (NML), obese (OC), obese-melatonin (OM), obese-leptin (OL), obese-melatonin-leptin (OML) groups. Body weights, peri-ovarian fat pads, volumetric parameters and numerical values of follicles were estimated. Also, the LH receptor (LHr) immune-positivity, catalase (CAT) and the myeloperoxidase (MPO) levels were determined. The body weight and peri-ovarian fat pads were significantly decreased following Mel (p < .05) treatment and, especially, Lep (p < .01) treatment. But, the ovarian weights were significantly increased following Lep (p < .05) and Mel (p < .01) treatment, in particular. The ovarian and cortex volume decreased in the OC group, and the cortex volume of the OC group was significantly higher than the Ob + Mel, Ob + Lep and Ob + Mel + Lep groups (p < .01). Besides, the volume of the cortex in the NL group was significantly higher than in the other groups (except for the NC group) (p < .01). Although, the total numbers of primordial and primary follicles in NC group were significantly higher than in the OC group (p < .001), the number of the primordial and primary follicles in OC group was significantly higher than in the OL (p < .05), OM (p < .05) and, especially, the OML groups (p < .001). Likewise, the number of the secondary follicles in the OML group was significantly less than that in the OC group (p < .05). The CAT and MPO activity of the OC group was significantly higher than in the NC group (p < .05) and also granulosa cell apoptosis had increased in obese rats; but it was decreased after Lep and Mel treatment. Otherwise, Lep and, in particular, Mel increased LHr positivity. We concluded that obesity could trigger abnormal ovarian function and polycystic ovary via inducing LHr apoptosis and suppressing ovarian folliculogenesis. Also, melatonin could be better for inhibition of apoptosis and modulation of folliculogenesis than leptin. These observations suggest that melatonin may act to reduce fertility in obese patients.Impact statementWhat is already known on this subject? Hormonal changes during reproductive cycle in obese women are particularly studied and there is not any study that evaluates the effects of melatonin and leptin, together.What the results of this study add? The study has shown that obese rats have increased granulosa cell apoptosis and MPO activities but melatonin and leptin reduces the apoptosis and inflammation. Moreover, the obesity decreased, but melatonin and leptin increased LHR immunoreactivity in both the granulosa and theca cells.What the implications are of these findings for clinical practice and/or further research? The results suggest that leptin and melatonin could decrease excess body weight in obese persons. Also, these hormones modulate the ovarian turn-over by regulating developing follicles. Therefore, leptin and especially melatonin could be used as a supplement to ovulation therapy.

Percutaneous Plantar Fascia Release With Needle: Anatomic Evaluation with Cadaveric Specimens.

Percutaneous plantar fascia release with needle is a novel procedure for the treatment of plantar fasciitis. The objective of this cadaveric study is to perform an anatomic evaluation of the percutaneous plantar fascia release method using a conventional hypodermic needle. In this study, we used 14 fresh-frozen cadaveric trans-tibial amputation specimens. Percutaneous plantar fasciotomy with a conventional hypodermic needle was performed. After a proper dissection, the width of the plantar fascia, the thickness of the medial border, and the width of the cut segment were measured and recorded. Any muscle damage on the flexor digitorum brevis and damaged area depth were recorded. Any damage on the lateral plantar nerve and the first branch of the lateral plantar nerve, also known as Baxter's nerve, and their distance to fasciotomy were also recorded. Mean width (± standard deviation) of the plantar fascia was measured as 20.34 ± 4.25 mm. The mean thickness of the medial border of the plantar fascia was 3.04 ± 0.54 mm. Partial fasciotomy was performed in all cadavers with 49.47% ± 7.25% relative width of the plantar fascia. No lateral plantar nerve, or its first branch Baxter's nerve, was damaged, and the mean distance from the deepest point of the fasciotomy up to the Baxter's nerve was 8.62 ± 2.62 mm. This cadaveric study demonstrated that partial plantar fasciotomy can be achieved via percutaneous plantar fascia release with a conventional hypodermic needle without any nerve damage.

Biomechanical and Morphometric Properties of the Long Flexor Tendons of the Toes: A Cadaver Study.

BACKGROUND: We sought to show the biomechanical and morphometric properties of flexor hallucis longus (FHL) and flexor digitorum longus (FDL) tendon grafts harvested by specific surgical approaches and to assess the contribution of FHL slips to the long flexor tendons of the toes. METHODS: Thirteen fresh-frozen amputated feet (average age, 79 years) were dissected. The connections between the FHL and FDL tendons and the contribution of FHL slips to the long flexor tendons were classified. The biomechanical properties of the tendons and slips were measured using a tensile device. RESULTS: The connections between the FHL and FDL tendons were reviewed in two groups. Group 1 had FHL slips (11 cases) and group 2 had cross-slips (two cases). The FHL slips joined the second and third toe long flexor tendon structures. Tendon length decreased significantly from the second to the fifth toe (P < .001). Apart from the second toe tendon being thicker than that of the fourth toe (P = .02) and Young's modulus being relatively smaller in the third versus the fourth toe tendon (P = .01), biomechanical and morphometric properties of second to fourth tendons were similar. Mechanical properties of those tendons were significantly different from fifth toe tendons and FHL slips. Morphometric and biomechanical properties of FHL slips were similar to those of the fifth toe tendon. CONCLUSIONS: Herein, FHL slips were shown to have biomechanical properties that might contribute to flexor functions of the toes. During the harvesting of tendon grafts from the FHL by minimally invasive incision techniques from the distal plane of the master knot of Henry, cutting slips between FHL and FDL tendons could be considered a cause of postoperative function loss in toes.

Comparing Dimensions of Four-Strand Hamstring Tendon Grafts with Native Anterior and Posterior Cruciate Ligaments.

Background. The aim of the study was to evaluate whether or not there was any incompatibility between four-strand hamstring tendons taken from the same knee and the dimensions of the ACL and PCL. Methods. 15 fresh frozen cadaver hamstrings were prepared as four-strand grafts and measurements made of the ACL and PCL circumferences in the midsection were made in the narrowest part of the midsection. The cross-section areas and diameters were calculated with geometric calculations used to measure the cross-sectional area of cylinders. Accepting that the geometric insertions were elliptical, the length, width, and area were calculated for entry areas. Results. A significant relationship at 96.2% was determined between the ACL mid and the hamstring diameter. A significant relationship at 96.7% was determined between the ACL and the hamstring mid area. A significant relationship at 96.4% was determined between the PCL mid and the hamstring diameter. A significant relationship at 95.7% was determined between the PCL and the hamstring mid area. Conclusion. For the reconstruction of ACL and PCL, it was determined that there is less incompatibility between the four-strand hamstring tendons taken from the same knee and the dimensions of the midsection PCL compared to the ACL dimensions.

The beneficial effects of Momordica charantia (bitter gourd) on wound healing of rabbit skin.

Momordica charantia (MC; bitter gourd) is a traditional herbal commonly used for its antidiabetic, antioxidant, contraceptive and antibacterial properties. In the current study, the authors aim to observe the topical effect of MC cream on the wound-healing process in rabbits. Moreover, they compare the healing potential with conventional creams used therapeutically. Towards this aim, 28 New Zealand rabbits were divided into four groups and excision wounds (7 cm²) were made on their backs. Open wound dressing was carried out daily for 28 days among the experimental groups with the application of dekspanthenol (Bepanthen®; BP group, n = 7), nitrofurazon (Furacin®; FR group, n = 7) and olive oil extract of MC (MC group, n = 7). No application was made to the control group. At the end of day 28, areas of the skin with initial wound area were en bloc dissected and prepared for histopathological and stereological analysis. Inflammatory cells were abundant in the control group and cream application led to a decrease in the number of these cells, especially in the MC group. The highest number of fibroblasts was detected in the MC group. Furthermore, the MC group displayed the highest fractions of epidermis to papillary dermis, fibroblasts to reticular dermis and collagen fibres to reticular dermis. The MC group also presented a high density of blood vessels, moderate density of collagen fibres and mature fibroblasts. The BP group showed better epithelialisation compared with the FR group, but the latter provided more effective reorganisation of the dermis. Different cream supplements caused healthy and fast wound healing according to untreated controls and the results show that administration of the MC extract improves and accelerates the process of wound healing in rabbits in comparison with the BP and FR extracts.