Comparative Study on Bone Marrow-Versus Adipose-Derived Stem Cells on Regeneration and Re-Innervation of Skeletal Muscle Injury in Wistar Rats

BACKGROUND@#Skeletal muscle injuries are frequent clinical challenges due to associated fibrosis and disability.Regenerative medicine is an emerging promising strategy for such cases. The aim of this study was to compare between the effects of bone marrow-mesenchymal stem cells (BM-MSCs) versus adipose tissue stromal cells (ADSCs) on regeneration and re-innervation of skeletal muscle laceration injury in Wistar rats at different time intervals. @*METHODS@#Six young male rats were used as a source of allogenic MSCs. Eighty-four adult female rats were divided into: Group I (control), Group II (Untreated Laceration): right gluteal muscle was lacerated and left for spontaneous healing, Group III (BM-MSCs): right gluteal muscle was lacerated with concomitant local intramuscular injection of 1 x 106 BM-MSCs in the lacerated muscle, Group IV (ADSCs): right gluteal muscle was lacerated with concomitant local intramuscular injection of 1 x 106 ADSCs in lacerated muscle. Rats were sacrificed after one, two and eight weeks.Muscles were processed to prepare sections stained with H&E, Mallory’s trichrome and immune-histochemical staining (neurofilament light chain). @*RESULTS@#A significant increase in collagen fibers and failure of re-innervation were noticed in untreated laceration group. BM-MSCs-treated groups showed regeneration of muscle fibers but with increased collagen fibers. Meanwhile, ADSCs showed better regenerative effects evidenced by significant increase in the number of myotubes and significant decrease in collagen deposition. Re-innervation was noticed in MSCs-injected muscles after 8 weeks of laceration. @*CONCLUSION@#Both BM-MSCs and ADSCs improved regeneration of skeletal muscle laceration injury at short- and long-term durations. However, fibrosis was less in ADSCs-treated rats. Effective re-innervation of injured muscles occurred only at the long-term duration.

Comparative Study on Bone Marrow-Versus Adipose-Derived Stem Cells on Regeneration and Re-Innervation of Skeletal Muscle Injury in Wistar Rats

BACKGROUND@#Skeletal muscle injuries are frequent clinical challenges due to associated fibrosis and disability.Regenerative medicine is an emerging promising strategy for such cases. The aim of this study was to compare between the effects of bone marrow-mesenchymal stem cells (BM-MSCs) versus adipose tissue stromal cells (ADSCs) on regeneration and re-innervation of skeletal muscle laceration injury in Wistar rats at different time intervals. @*METHODS@#Six young male rats were used as a source of allogenic MSCs. Eighty-four adult female rats were divided into: Group I (control), Group II (Untreated Laceration): right gluteal muscle was lacerated and left for spontaneous healing, Group III (BM-MSCs): right gluteal muscle was lacerated with concomitant local intramuscular injection of 1 x 106 BM-MSCs in the lacerated muscle, Group IV (ADSCs): right gluteal muscle was lacerated with concomitant local intramuscular injection of 1 x 106 ADSCs in lacerated muscle. Rats were sacrificed after one, two and eight weeks.Muscles were processed to prepare sections stained with H&E, Mallory’s trichrome and immune-histochemical staining (neurofilament light chain). @*RESULTS@#A significant increase in collagen fibers and failure of re-innervation were noticed in untreated laceration group. BM-MSCs-treated groups showed regeneration of muscle fibers but with increased collagen fibers. Meanwhile, ADSCs showed better regenerative effects evidenced by significant increase in the number of myotubes and significant decrease in collagen deposition. Re-innervation was noticed in MSCs-injected muscles after 8 weeks of laceration. @*CONCLUSION@#Both BM-MSCs and ADSCs improved regeneration of skeletal muscle laceration injury at short- and long-term durations. However, fibrosis was less in ADSCs-treated rats. Effective re-innervation of injured muscles occurred only at the long-term duration.

effect of bone marrow-derived mesenchymal stem cells on the healing experimental skin wound in diabetic adult male albino rats

Impairment of wound healing in diabetic patients is an important clinical problem. Various studies have suggested that using regenerative medical therapy could be a good solution. Mesenchymal stem cells are nonhematopoietic progenitor cells that have high differentiation potential and could be isolated from bone marrow and other tissues. The aim of this study was to assess the role of stem cells in the healing of diabetic skin wounds of male albino rats. Forty-two male albino rats were used in this study. Seven of them of an average weight of 50-70 g were used for extraction of bone marrow. The rest of the rats were of an average weight of 150-200 g and were used as animal models. The animals were divided into four groups: group I, comprising rats from which the bone marrow was extracted; group II, which formed the control group; group III, comprising diabetic wounded rats; and group IV, in which wounded diabetic rats were treated with bone marrow-derived mesenchymal stem cells. Diabetes was induced in rats by means of a single intraperitoneal injection of streptozotocin. Wounds of groups III and IV were examined after 3 and 7 days. Skin specimens were processed and stained with H and E and Masson's trichrome. Re-epithelization with complete closure of the wound was noted in the stem cell-treated group after 7 days. There was a significant increase in the number of hair follicles/high-power field and increase in the collagen content of the dermis in the stem cell-treated groups compared with the untreated groups. Bone marrow-derived mesenchymal stem cells can be effectively used in the treatment of diabetic wounds

Prevalence of venous thrombo-embolism in acute exacerbations of chronic obstructive pulmonary disease

Objectives: To determine the prevalence of venous thrombo-embolism in patients with acute exacerbations of COPD

Subjects and methods: This was a cross-sectional study involving 105 patients hospitalized with exacerbations of COPD. Clinical and hematological parameters on admission were collected. Multislice computed tomographic pulmonary angiography [CTPA] and ultrasonographic examination of lower limbs, for pulmonary embolism and deep vein thrombosis respectively were done. Wells and Geneva scores were calculated

Results: This study was conducted on 105 COPD patients with acute exacerbations. All of them were males with mean age 49.3 + 8.43. Pulmonary embolism was found in 28.6% of COPD patients who were definitely diagnosed by CTPA while DVT was found in 26.7% of positive cases of pulmonary embolism detected by venous duplex. Wells and Geneva scores were calculated, high probability Wells score was found in 83.3% while Geneva score was likely in 90% of COPD patients proved to have pulmonary embolism. D-dimer and CBC were done; D-dimer was found negative in 90.0% in patients proven not to have pulmonary embolism while was positive in 100% of the diagnosed patients. Regarding CBC; polcythemia was found in 73.3% of diagnosed cases

Conclusion: VTE appeared to be a common problem in COPD patients with exacerbations. The role of CTPA is the cornerstone in the diagnosis of pulmonary embolism. DVT of lower limbs was not essential in all cases of proven pulmonary embolism. Serum D-dimer, Wells criteria and Geneva score are useful bedside criteria that may help to assess the occurrence of VTE in such patients

[Identification and evaluation of the probiotic effect of isolated lactic acid bacteria in traditional goat cheese]

In order to identify lactic acid bacteria [LAB] associated with traditional goat cheeses and to assess their probiotic effect, chemical and microbiological analysis were carried out on seven products collected from different Lebanese regions: Laban [L], Labneh Ambarise[LA], Labneh Serdalli [LS], Jebneh Darfieh [JD], Keshek [K], Shanklish [Sh] and Labneh Darfiyeh [LD]. These samples were analysed for their pH and acidity. The isolation of lactic acid bacteria was achieved using two selective media, namely MRS and M17 agar. Isolated strains were identified according to morphology, Gram staining, catalase and oxydase, ammonia production from arginine, carbon dioxide production from glucose, growth in MRS broth at 10degree C and 45degree C, salt tolerance [2, 4 and 6.5% of NaCl], sugar fermentation with API [Analytical Profile Index] system methods [50CHL and 20Strep]. The probiotic potential of isolates to inhibit pathogenic bacteria [Salmonella, Staphylococcus aureus and E. coli], their survival at different pH [1, 2, 3 and 7.2], the time needed to reach acidification end point in full and skimmed milk were assessed. Labneh Ambarise and Labneh Darfiyeh had an average pH of 4+/-0.5 and a titratable acidity of 2%+/-0.1 [w/w], while the average pH and titrable acidity of Darfiyeh cheese were 5.22 and 0.62 respectively. The presence of LAB in one year old samples preserved in olive oil could not be detected. Twenty five strains were isolated and identified from different samples, where nine of them belong to Lactobacillus plantarum [L2, Sh2, K2, K3, K4, K5, LS1, LS3 and LS4], three are Lactobacillus paracasei spp. paracasei [Sh3, LS2 and LS5] and five are Leuconostoc mesenteroides spp. dextranicum 2 [JD1, JD2, JD3, JD4 and JD5]; four colonies are suspected to be Lactobacillus casei [L1, Sh4, K1 and K6] and four others are probably yeasts [L3, L4, L5 and L6]. Identified strains showed a capacity to produce antimicrobial substances with maximum inhibition diameters of 20 mm against Salmonella [K1, Sh4, LS1 and JD1], 21 mm and 19 mm for L2 against Escherichia coli and Staphylococcus aureus, respectively. The microorganisms survival rate in simulated gastric juice [SGJ] varied between 50% and 93% at pH 3 [Sh3, K2, LS2, LS5, JD1 and JD3]. Their growth was inhibited at pH 1 [0.05% to 0.08%]; only JD1 and JD3 showed more then 50% of viability at pH 2. The time needed for milk acidification had an average of more than 10 h exceeding the time required by yogurt starter cultures [5 h]. The Jebneh Darfieh sample, rich in Leuconostoc mesenteroides, showed the most interesting characteristics of cell viability at pH 2 [50% for JD1 and JD2] and a maximum of 20 mm of inhibition against Salmonella. It could be used as a natural source of probiotic. The isolates of the samples Laban, Shanklish, Keshek and Labneh Serdalli [L. plantarum and L. paracasei mainly] were more sensitive to high acidity [pH=2], but showed good antimicrobial activity against pathogens. Microencapsulation is probably required to protect them from harsh conditions. Further tests are needed to determine the exact identity of some strains and the nature of secreted substances responsible of the probiotic effects

Screening for minimal hepatic encephalopathy in asymptomatic drivers with liver cirrhosis

Minimal hepatic encephalopathy [MHE] represents a part of the spectrum of hepatic encephalopathy [HE]. It can have a far-reaching impact on quality and ability to function in daily life and may progress to overt HE. This study was designed to screen for MHE in drivers with liver cirrhosis in Mansoura, a city in the Nile delta in Egypt. A total of 174 consecutive drivers with positive serology for viral markers and cirrhosis were screened for MHE. Questionnaires and standard psychometric tests and well-informed consent were performed at the same setting. The diagnosis of MHE was made when one or both symbol digit test [SDT] and number connection test [NCT] appeared abnormal. Beck's inventory and Mini Mental State Examination questionnaires were performed for those diagnosed as MHE. After overnight fasting, venous blood samples were taken for haematologic tests and routine liver function tests by conventional methods. Arterial ammonia was also measured. A total of 66 patients showed evidence for MHE out of 139 patients who fulfilled the inclusion criteria. No significant differences were present, apart from a significantly elevated arterial ammonia level [p-value <0.001] and a bad self-reported driving history [p < 0.05] in the MHE-positive group when compared with the MHE-negative group. Multivariate logistic regression revealed that advanced Child-Pugh grade [p < 0.001], hepatitis B virus [HBV]-related a etiology [p < 0.001] and smoking are significant risk factors for MHE. MHE is significantly commoner among Child-Pugh C patients [p < 0.05] when compared with the other Child-Pugh grades. Our data revealed a high prevalence of MHE [47%] among Egyptian drivers with liver cirrhosis. It is hence recommended to include the driving history as well as regular pencil-paper standard psychometric testing in evaluating those at risk, especially in the outpatient setting, for early detection and proper management

comparative study of the static histomorphometry and marrow content of human vertebral and iliac crest trabecular bone

Cancellous bone architecture is an important determinant of bone strength. The iliac crest marrow is a gold standard graft during discectomy and spinal fusion procedures but is associated with donor site morbidity. In a trial to find an alternative, the aim of this work was to compare the structure and histomorphometric measurements of the vertebral and iliac bones and assess their bone marrow cellular content and osteogenic potential. Vertebral body bone specimens were taken from four human subjects and specimens of iliac crest bone were obtained from five human subjects scheduled for surgery after taking their consent. Histomorphometric measurements were done in every specimen using decalcified and undecalcified sections. Osteogenic potential of the marrow progenitor cells was estimated by immunohistochemical detection of alkaline phosphatase expression. Both vertebral body and iliac crest were seen consisting of branching and anastomosing trabeculae covered by cortical bone. However, the trabecular thickness and trabecular separation were significantly higher in the vertebral body than in the iliac crest. Moreover, the osteoid thickness and eroded surface were significantly greater in the vertebral body indicating more turning over activity as compared to iliac crest. The marrow content of nucleated cells was highly significant higher in vertebral body than that of the iliac crest. In addition, the osteogenic potential of the marrow progenitor cells showed no significant difference between vertebral body marrow and that of iliac crest. Static histomorphometric measurements of vertebral body differ from that of iliac crest. Meanwhile, vertebral bone marrow is rich in progenitor cells and its osteogenic potential is nearly the same as that of iliac crest, so vertebral marrow could be considered a suitable candidate for marrow graft supplementation in spinal fusion procedures

Histological study of the role of medical ozone in modulating experimentally induced chronic liver injury in male albino rats

Medical ozone, a mixture gas of ozone and oxygen is used as a therapy for various diseases and has been recently used in viral hepatitis. This study investigated the possibility that ozone treatment may decrease or prevent the development of liver cirrhosis in chronic liver diseases using rats taking carbon tetrachloride. 35 rats were used and were divided into 4 groups. Rats of group I act as control and rats of group II were given medical ozone. Carbon tetrachloride was given to rats of group III while rats of group IV were given both carbon tetrachloride and medical ozone on alternating days. Chronic administration of carbon tetrachloride resulted in degeneration of most hepatocytes together with disorganization of their arrangement. There was a significant increase in the number of smooth muscle actin positive cells/hpf and in the collagen content in the portal tract areas. The fenestrations of the endothelial cells lining the sinusoids were rarely seen and basement membrane material appeared under the sinusoidal wall. Co-administration of medical ozone with carbon tetrachloride to rats preserved the integrity of most hepatocytes and preserved their content of the rough endoplasmic reticulum. However, small vacuoles were still seen in the hepatocytes and there was still disorganization of the hepatocyte arrangement. The collagen content was significantly decreased as compared to group III but still showed highly significant increase as compared to control. Medical ozone can preserve the integrity of many hepatocytes in chronic liver injury and has a moderate effect on the collagen fiber formation. So, it is recommended to do clinical trials to investigate the ability of medical ozone to preserve the liver functions in chronic liver diseases

Histological and scanning electron microscopic study of the effect of cigarette smoke on the heart of male albino rats

Environmental tobacco smoke has been shown to cause heart diseases among non-smokers. Twenty one adult male albino rats were used to investigate the effect of cigarette smoke on the heart and to evaluate the possibility of recovery. Rats were divided into 3 equal groups. Rats of group I were considered as control Rats of group II were exposed to cigarette smoke for 4 weeks, while those of group III were exposed to cigarette smoke for 4 weeks then the exposure was stopped for another 4 weeks. Exposure to cigarette smoke resulted in affection of the coronaries. Some cardiac muscle fibers showed vacuolated cytoplasm, while other showed deeply stained acidophilic cytoplasm and psychotic nuclei. Scanning electron microscopic examination showed that some cardiac muscle fibers appeared necrotic with loss of their normal architecture. There was a decrease in succinic dehydrogenase enzyme activity and in the antiapoptotic Bcl-x [L]protein expression. Stopping the exposure for 4 weeks showed that few cardiac muscle fibers were still affected. Areas of fibrosis were seen between the cardiac muscle fibers in Mallroy stained sections and in the scanning electron microscopic study. There was an increase in scenic dehydrogenase enzyme activity and in the Bcl-x[L] protein expression in the cardiac muscle fibers in group III- that was left for 4 weeks for recovery after 4 weeks of exposure to cigarette smoke- in comparison to those of group II, but was still less than those of the control It is concluded that cigarette smoke affected the cardiac muscle and some of its effects could not be reversible

role of angiotensin converting enzyme inhibitors on the prevention of myocardial necrosis in experimental animals

The present work is concerned with the study of the role of the angiotensin converting enzyme inhibitors [A. C. E. Is.], enalapril and captopril, in the preservation of the myocardium in isoprenaline-induced ischemic changes in rats. It was found that capftpril in a single dose could significantly reduce the percentage of ischemic changes. On the contrary, enalapril was insignificantly effective. On the other hand subacute [15 days] administration of enalapril could significantly reduce the percentage of myocardial necrosis, more than captopril in animals treated with the drug for the same period. It was concluded that the variability in the pharmacokinetics and hemodynamic effects of both drugs is the cause of difference in the response to each drug. Also,it was concluded that the A. C. E. Is. could be effective in treatment of hypertension with ischemic heart disease, through their coronary dilator effect, through inhibition of the angiotensin-renin system, and also, by the lack of unwanted effects on lipid metabolism

Reactivity centres among nucleophilic substituted arylidene acetophenone towards attack by active methylene group [Part XI] synthesis of newer carboxylic acids, cyclic hydrazides, isoxazole and pyridine derive

Hans stobbe [1] stated that alpha beta-unsaturated ketones failed to undergo the Stobbe condensation but form Michael adducts. Patwardhan [2] generalized the reactivity of the carbonyl group in the presence of potassium tert-butoxide, giving the corresponding half-esters in a low yield. Recently [3], we have reported on the reactivity of both centers with special attention to the participation in the Michael and Stobbe condensation. The results obtained in the present study are considered to be rather conclusive in establishing the role of the polar factor in the reactivity of the carbonyl carbon in chalkones. To undergo the Stobbe condensation in a good yield, there must be an electron donating substituents which favours the formation of the intermediate lactone proposed in the currently accepted mechanism[4]] Secondly there must be high concentration of ester group for consuming the protonium ion in alcohol formation. The protonium ion assumed as a by-product [5] and leading to the formation of the competing Michael addition at the expense of the Stobbe condensation. The present investigation was planned to substantiate this conclusion. Thus twelve chalkones having substituents of different polar nature were condensed with succinic ester in the presence of potassium tert-butoxide and the composition of the reaction products were determined. The results obtained may be considered as a contribution characterized to the work done by Hans Stobbe and Patwardhan. An elegant support for the importance of the polar factor can be shown by the failure of the alpha, beta-unsaturated ketones having an electron attracting group to undergo the Stobbe condensation, where the maximum yield range from 3-10 percent it is interesting to note that the yields of the half-esters increase with the increase of the number of the methyl and methoxyl substituents, whereby the half-ester attains 70 percent when the tetra-substituted chalkone was utilized in the Stobbe condensation. For this purpose we reacted 4,4'- dimethoxy, 4-methoxy 3/4'-dimethyl and 2.3-dimethoxy 3', 4'-dimethyl chalkones with dimethyl succinate to verify good yields of the half-esters [see Experimental]. The IR spectra of these compounds showed absorption bands of the dimeric carboxylic groups for esters and acids. Alkaline hydrolysis of the half-esters gave the corresponding dibasic acids of the type [Ib] and [IIb]. The structure of these acids was supported by analytical data, IR spectra and the facile conversion into the corresponding cyclic hydrazide [IIIa],upon treatment with hydrazine hydrate. The IR spectrum of compound IIIa showed characteristic absorption bands at 3380, 1660, 1620 and 1600 cm[-1] The reactions with dimethyl sulphate [6] and methyl isocyanate to form dimethyl and dicarba-mate succinhydrazide derivatives, IIIb, c were considered as additional evidence for the assigned structure IIIa The reaction of chalkones and cyanoacetamide in the presence of ammonium acetate afforded cyanopyridones [IVa],[IVb] and cyanodihydropyridone [V] [7], rested upon correct analytical and spectral data. When 4-methoxy 3',4'-dimethyl chalkone was brominated in acetic acid, it afforded the dibromide which upon treatment with hydroxylamine hydro-chloride under conditions similar to those described by Nakata [8] for isoxazole synthesis, compound VI was obtained, the structure of which is supported by a study of electronic spectrum which is similar to that previously described [9]. It has been assumed [10] that the isoxazoles are obtained through the formation of acetylenic oxime presumably formed as an intermediate through dehydrobromination, followed by intramolecular cycloaddition. The reaction of 4-methoxy 3,'4'-dimethyl chalkone with ethylcyanoacetate in presence of ammonium acetate, afforded VII. Whether the reaction was carried out with or without a solvent, the same product was obtained. However, compound VII is not identical with that formed by Sakurai and Midorikawa [11] for the reaction of unsubstituted alpha, beta- unsaturated ketones with ethylcyanoacetate as shown by spectral and analytical data. The reaction of 4-methoxy 3', 4 -dimethyl chalkone with malononitrile [12] afforded cyanoaminopyridine [VIII]. The IR spectrum showed the frequency bands due to asymmetric and symmetric stretching vibrations of primary amines as multiple bands of medium intensity. The corresponding bending vibrations cause absorption in the 1630 cm[-1] region as 2-amino pyridine, in addition to the characteristic absorption bands for the cyano group

Heterocycles from dicarboxylic acids Synthesis of new lactones, succinhydrazides and pyridazine derivative XIII

The title compounds were prepared via two different routes. Thus the dicarboxylic acids [I] and [II] previously prepared by the author [1], when treated with bromine in acetic acid, lactones [IIIa, b] were formed through the elimination of hydrogen bromide [2]. The I.R. spectrum of these compounds showed the characteristic absorption bands for lactones and carboxylic acids

Reactivity centres in the dimethoxybenzylidene acetophenone towards attack by active methylene compounds [Part X]: synthesis of five membered ring compounds

While malononitrile and cyanoacetamide react with 2,3-dimethox-y benzylideneacetophenone to give pyridine and pyridone derivatives, through Michael condensation, the reaction with succinic ester condenses in the Stobbe type affording the half-ester [IIa]. This undergoes cyclization to five membered ring compounds via two different routes. The synthesis of isoxazole [IX], oxadiazole [XIa] and thiazole [Xlb] is also described. The biological activity of these compounds is evaluated and discussed

Heterocycles and oxime carbamates from arylidene indanones Reactions of indanone with aldehydes and succinic esters: Part [XIV]

It has been stated [1] that the condensation of aldehydes with indanone gave the corresponding arylidene indanone derivatives