Comparative characteristics of mesenchymal stem cells derived from reamer-irrigator-aspirator, iliac crest bone marrow, and adipose tissue.

Mesenchymal stem cells (MSCs) have been considered promising tools for new clinical concepts in supporting cellular therapy and regenerative medicine. More recently, Ream/Irrigator/Aspirator (RIA) was introduced as a source of MSCs. In this study we compared MSCs derived from three different sources (iliac crest bone marrow (ICBM), adipose tissue (AT), and (RIA)) regarding the morphology, the success rate of isolating MSCs, colony frequency, expansion potential, osteogenic and chondrogenic differentiation capacity. MSCs were isolated from three different sources and flow cytometric analyses were performed for cell characterization. Colony-forming unit-fibroblast (CFU-F) assay and population doubling time (PDT) were evaluated for MSCs derived from three different sources and differentiation potential of RIA, ICBM-, and AT-MSCs were determined by staining. Additionally, gene expression profiles for tissue specific markers corresponding to osteogenesis and chondrogenesis were analyzed using real time polymerase chain reaction (RT-PCR). Cultured with the appropriate condition, osteogenic and chondrogenic differentiation could be confirmed in all MSC preparations. Flow cytometry analysis indicated that RIA- and AT-derived MSCs have more homogenous populations than ICBM-MSCs. A comparison of the colonogenic ability in different tissues by CFU-F assay after 10 days showed that more colonies are formed from RIA-MSCs than from ICBM-MSCs, and AT-MSCs. AT-MSCs, were dispersed with no obvious colonies. The RIA-MSCs underwent osteogenesis and chondrogenesis at a faster rate than ICBM and AT-MSCs. Direct comparisons of RIA- to ICBM- and AT-MSCs have shown the RIA-MSCs have higher differentiation toward osteoblast and chondrocytes compared to other sources of MSCs. Hence, RIA-MSCs may be recommended as a more suitable source for treating orthopedic disorders.

Hypoxia and laser enhance expression of SDF-1 in muscles cells.

Targeted homing of transplanted mesenchymal stem cells (MSCs) is a decades old discussion in regenerative medicine. It has been proved that stromal cell-derived factor-1 (SDF-1α) is a potent chemoattractant of MSCs. Therefore, different strategies have been used to increase secretion of SDF-1α in damaged tissues to elevate targeted homing of MSCs. Previous studies have revealed that increased SDF-1α expression in hypoxic necrotic tissues and also low-level laser exposure enhanced angiogenesis in injured tissues. Herein, human skeletal and cardiac muscle cells (HSKM and HCM) were treated with hypoxia and low level laser to see their effects on expression of SDF-1α and on MSCs migration towards these treated cells. The optimal treatment conditions were determined by investigating the cellular viability after treatment. Real-Time PCR and Western blot analysis were done to study the expression of SDF-1α in treated cells. Migration potential of MSCs toward hypoxic and laser treated cells was investigated via migration assay. MTT assay revealed that laser and hypoxia treatment had no effect on the viability of HCM, HSKM compared with Glioblastoma cells. Real-Time PCR showed 16- and 90-fold elevation in mRNA of SDF-1α in HSKM and HCM cells, respectively, in laser treated with 12 J/cm2 intensity. In these two groups, selected as optimal conditions, HIF-1α expression showed maximum fold changes that might be partly because of response to treatments help to SDF-1α expression. It can be concluded that hypoxia and laser treatments may recruit MSCs and applied as a useful strategy for the further targeted stem cell homing.

Adipose derived mesenchymal stem cells express keratinocyte lineage markers in a co-culture model.

Cutaneous wound healing is a complex type of biological event involving proliferation, differentiation, reprograming, trans/de-differentiation, recruitment, migration, and apoptosis of a number of cells (keratinocytes, fibroblasts, endothelial cells, nerve cells and stem cells) to regenerate a multi-layered tissue that is damaged by either internal or external factors. The exact regeneration mechanism of damaged skin is still unknown but the epithelial and other kinds of stem cells located in skin play crucial roles in the healing process. In this work, a co-culture model composed of adipose derived mesenchymal stem cells and keratinocytes was developed to understand the cellular differentiation behaviour in wound healing. Human mesenchymal stem cells were isolated from waste lipoaspirates. Keratinocytes were isolated from neonatal rats skin as well from human adult skin. Both types of cells were cultured and their culturing behaviour was observed microscopically under regular intervals of time. The identity of both cells was confirmed by flow cytometry and qRT-PCR. Cells were co-cultured under the proposed co-culturing model and the model was observed for 7, 14 and 21 days. The cellular behaviour was studied based on change in morphology, colonization, stratification, migration and expression of molecular markers. Expression of molecular markers was studied at transcriptional level and change in cellular morphology and migration capabilities was observed under the invert microscope regularly. Successfully isolated and characterized mesenchymal stem cells were found to express keratinocyte lineage markers i.e. K5, K10, K14, K18, K19 and Involucrin when co-cultured with keratinocytes after 14 and 21 days. Their expression was found to increase by increasing the time span of cell culturing. The keratinocyte colonies started to disappear after 10 days of culturing which might be due to stratification process initiated by possibly transdifferentiated stem cells. It can be concluded that mesenchymal stem cells can regenerate the damaged skin if transplanted to damaged area but for their successful differentiation and enhanced regeneration, they need a population of keratinocytes in situ which need further experiments for validation of co-culture model and its potential for being used in clinics.

Pancreatic pathology in chronic dialysis patients--an autopsy study of 78 cases.

Autopsy data from 78 patients with end-stage renal disease (ESRD) treated with long-term hemodialysis were examined. Various pancreatic abnormalities were found in 47 (60%) patients. The most prevalent abnormality was pancreatitis which was seen in 22 patients (28%). Other lesions found with considerable frequency included fibrosis, hemosiderin deposits, calcification, cystic changes, amyloidosis and abscess formation. In addition hyalinization, atrophy or absence of islands of Langerhans and necrosis of peripancreatic fat were seen in several cases and inspissated secretions, focal ductular epithelial metaplasia and dilatation were noted in some patients. Comparison of the present data with those of a large survey of ESRD patients conducted prior to dialysis era indicates a considerable increase in the prevalence of pancreatic pathology in ESRD patients sustained by long-term hemodialysis treatment.

Hepatobiliary pathology in hemodialysis patients: an autopsy study of 78 cases.

Necropsy findings of hepatobiliary system from 78 patients with end-stage renal disease maintained on hemodialysis are reported. Ninety percent of the patients exhibited some abnormalities. Multiple abnormalities often coexisted in each patient. Hepatomegaly was found in 50% of the patients and could be attributed to a discernible cause in all but two of the affected patients who had isolated hepatomegaly. Hepatic congestion was also prevalent and was complicated by fibrosis, cardiac cirrhosis, and centrilobular necrosis and hemorrhage in some patients. This was associated with chronic fluid overload, hypertension, and/or cardiovascular disease in the affected patients indicating the importance of adequate control of these factors. Mild periportal hepatic fibrosis, fatty metamorphosis, triaditis, hemosiderosis, and cystic changes were also seen with some frequency--the latter were associated with polycystic kidney disease and were complicated by massive intracystic hemorrhage and abscess formation, each in one patient. Chronic active hepatitis was found in three patients and was associated with chronic HBs antigenemia in one patient and presumed non-A, non-B infection in two. Nearly 22% of the patients showed either cholelithiasis at autopsy or before cholecystectomy due to complications. Significant negative findings included lack of acute viral hepatitis, silicone hepatosis, and recently described focal anoxic lesions associated with erythrocyte sludging. In conclusion, the present study has demonstrated the spectrum of hepatobiliary pathology in a large group of patients with end-stage renal disease maintained on hemodialysis.

Pathology of gastrointestinal tract in chronic hemodialysis patients: an autopsy study of 78 cases.

The pathological data obtained from postmortem examination of the gastrointestinal tract in 78 chronic hemodialysis patients are reported. All but two of the patients exhibited some gastrointestinal abnormalities. Multiple abnormalities often coexisted in one individual. Esophagitis, gastritis, duodenitis, enteritis, and colitis were seen with considerable frequency. However, extensive hemorrhagic, ulcerative, and pseudomembranous lesions frequently seen in untreated uremia were generally lacking. Evidence of peptic ulcer disease was found in one-fourth of the patients confirming the increased predisposition of dialysis patients to this complication. Several patients exhibited various ischemic phenomena including thrombosis, embolism, and infarction of various segments of the gastrointestinal tract. In addition, numerous other abnormalities were found throughout the gastrointestinal tract with lesser frequency. The data indicate that various gastrointestinal abnormalities commonly occur in patients with endstage renal disease despite uremia control by hemodialysis. However, the observed abnormalities are different from those reported in untreated uremia.

Features of residual renal function in end-stage renal failure associated with spinal cord injury.

The characteristics of end-stage renal disease (ESRD) complicating spinal cord injury (SCI) were studied retrospectively in 43 male hemodialysis patients. A control group of male patients dialyzed in the same institution were studied for comparison. The SCI patients had significantly lower serum creatinine concentrations and daily urinary creatinine excretion than the control group, despite comparable creatinine clearances. Therefore, serum creatinine, when compared with the familiar values in non-SCI patients, may greatly underestimate the severity of the renal impairment. Urine output was higher, urine specific gravity lower, and renal glucosuria more common in the SCI patients. 24-hour urinary protein excretion was higher and serum albumin was lower in the SCI patients, with 48% of the patients exhibiting nephrotic range proteinuria. Urine pH was markedly elevated, and pyuria and bacteriuria were present in all SCI patients. Fractional excretion of potassium (159 +/- 16%) exceeded its filtered load in most SCI patients.

Pathology of endocrine organs in chronic renal failure--an autopsy analysis of 66 patients.

Gross and microscopic findings at autopsy are reported in 66 patients with end-stage renal disease (ESRD). Nineteen patients received renal transplant, and 47 were maintained on chronic dialysis. Thyromegaly was seen in over half of the dialysis patients and in a third of the transplant recipients. The mean weight of the thyroid gland in the two groups was similar (26.2 +/- 4.5 g in the transplant group and 27.2 +/- 6 g in the group on dialysis). The parathyroid glands were hyperplastic in 10/11 dialysis patients (91%), but of normal size in 78% of the transplant recipients. Adrenal gland atrophy was common in the transplant group, probably reflecting adrenal suppression by glucocorticoids. Seminiferous tubular atrophy was observed in 12/16 dialysis patients and small testes were noted on gross examination in 50% of the transplant recipients. Prominent autopsy findings were goiter, parathyroid gland hyperplasia, and testicular atrophy. Following transplantation parathyroid gland hyperplasia declined, whereas goiter and testicular atrophy appeared to persist.

Erythropoiesis associated with viral hepatitis in end stage renal disease.

We describe an end stage renal disease (ESRD) patient on maintenance hemodialysis who developed a spontaneous increase in erythropoiesis associated with an episode of viral hepatitis. Resolution of the hepatitis was accompanied by a reduction in erythropoiesis with the hemoglobin and hematocrit falling back toward the patient's low baseline levels. Plasma erythropoietin (Ep) titers were measured during the period of active erythropoiesis and were found to be low to low normal. The unusual phenomenon of increased erythropoiesis in ESRD following liver injury has been previously described and is thought to be mediated through increased hepatitic Ep production. The low Ep titers measured in our patient, however, suggest that the liver may be capable of stimulating erythropoiesis by another mechanism.

Nutritional evaluation of hemodialysis patients with and without spinal cord injury.

Both end stage renal disease (ESRD) and spinal cord injury (SCI) represent major disabling conditions that may be associated with protein-calorie malnutrition (PCM). The prevalence of PCM in ESRD, however, remains unknown, and virtually no data exist regarding the status of PCM in patients with both SCI and ESRD. In this study we evaluated 23 ambulatory-ESRD patients and 11 SCI-ESRD patients utilizing a range of parameters recommended for assessing nutritional status in ESRD. Based on these parameters, our results show evidence for PCM in a substantial proportion of both groups. When the two groups were compared, however, the frequency and severity of PCM were significantly greater in the SCI-ESRD patients. Important factors felt to adversely influence nutritional status in the SCI-ESRD patients were intercurrent infection and amyloidosis that were found to frequently complicate this setting. It is also probable that the combined effects of PCM and ESRD significantly predispose these patients to further infection, establishing a vicious cycle.

Prediction of creatinine clearance from serum creatinine in spinal cord injury patients.

Measured endogenous creatinine clearance (Ccr) was compared with the predicted Ccr in 22 paraplegic, 36 tetraplegic and 11 ambulatory male individuals as well as 11 ambulatory females all of whom had normal renal function. While the predicted and measured values closely matched in the ambulatory patients the predicted values in the spinal cord injured patients consistently exceeded the measured values. It thus appears that the original Cockcroft and Gault formula; (formula; see text) when applied to SCI patients can be misleading. Modification of the original formula using a correction factor of 0.8 in paraplegics and 0.6 in tetraplegics was found to allow prediction of Ccr from age, sex, body weight, and serum creatinine in these patients with reasonable accuracy.

Clinicopathological characteristics of dialysis patients with spinal cord injury.

Forty-three spinal cord injured patients with endstage renal disease (ESRD) maintained on hemodialysis were studied. The most prevalent renal lesions consisted of chronic pyelonephritis and amyloidosis while the main renal functional features included nephrotic range proteinuria, high urine output and relatively low serum creatinine for the degree of renal insufficiency. Normocytic, normochromic anemia with low reticulocyte response, low serum iron and iron binding capacity and high transfusion requirement and serum ferritin were noted. Various cardiovascular, pulmonary and gastrointestinal abnormalities were found with considerable frequencies. The incidence of amyloidosis was much higher than that reported previously. This is thought to be due to continued progression of amyloidosis occasioned by longer survival in the present series.

Lung pathology in chronic hemodialysis patients.

To determine the spectrum and the relative frequencies of different pulmonary diseases in chronic renal failure (CFR) patients, we reviewed autopsy records from 46 chronic hemodialysis patients with end-stage renal disease of various etiologies. Pathological abnormalities were observed in the lungs of all but one patient. Acute and chronic lung diseases were found in 95.7% and 80.4% of subjects respectively. The most common acute diseases were pulmonary infections (pneumonia, lung abscess, empyema) and fluid overload. The most common chronic process was interstitial fibrosis. Other relatively common chronic diseases included pleural fibrosis and/or pleuritis as well as pulmonary arteriosclerosis, hemorrhage, thromboembolism and calcification. The incidences of granulomatous lung diseases, pulmonary amyloidosis and metastatic lung tumors were also higher than expected. Relative incidences of the different pulmonary diseases did not appear to be related to the etiology of CFR; rather, lung abnormalities were common in CFR of all etiologies.

Survival on maintenance dialysis in patients with chronic renal failure associated with paraplegia and quadriplegia.

Forty men with end-stage renal failure associated with spinal cord injury were treated with maintenance hemodialysis. There were 28 paraplegic and 12 quadriplegic patients. Survival on dialysis was significantly shorter in quadriplegics (5.5 +/- 5.0 months) than paraplegics (22.9 +/- 27.0 months). The cumulative I- and 2-year survival in paraplegics were 60 per cent and 52 per cent respectively. The respective values for quadriplegics were 34 per cent and 17 per cent. Various infections proved to be the immediate cause of death in the majority of our patients.

Hematologic features of chronic renal failure associated with spinal cord injury.

We examined the available hematologic data on 43 spinal cord injury (SCI) patients with end-stage renal disease (ESRD) treated with hemodialysis since 1972. A control group consisting of 45 ESRD patients without SCI was included for comparison. Moderate to severe anemia was found in all patients. The observed anemia was generally normocytic, normochromic, and associated with a low reticulocyte response. Transfusion requirement in the SCI group was considerably more than in the control group. There was a high incidence of splenomegaly, splenic amyloidosis, and lymph node enlargement in the SCI group. Lymph node and bone marrow amyloid deposits were found in some SCI patients but not in the control subjects.

Effects of long-term testosterone administration on pituitary-testicular axis in end-stage renal failure.

The endocrine effects of long-term testosterone administration were studied in 6 end-stage renal failure patients. During a 3-month control period where no androgens were administered the mean plasma testosterone level (7.3 nmol/l) was depressed while mean plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) levels were elevated at 41.2 mU/ml, 105.5 mU/ml, and 63 ng/ml, respectively. These values were repeated during a 6-month study period where each subject was administered testosterone enanthate (400 mg) intramuscularly once a week. Plasma testosterone levels markedly increased in all subjects with a mean elevation of 72.4 nmol/l, while reductions were observed in FSH and LH levels with values of 2.7 and 16.3 mU/ml, respectively. When compared with control period values, these changes were statistically significant (p less than 0.05). Although the mean plasma PRL level of 49.0 ng/ml was reduced when compared with the control period values, this reduction was not statistically significant. Our control period findings of low plasma testosterone levels coupled with high plasma LH and FSH are consistent with Leydig cell dysfunction. The significant reductions in plasma FSH and LH noted during the study period indicate a negative feedback effect produced by the pharmacologic doses of testosterone. Long-term testosterone administration, however, did not significantly affect the elevated mean PRL levels observed in these subjects.

Urolithiasis in patients with spinal cord injury.

The composition of the stone was determined in 24 paraplegic patients from whom 26 stones were surgically removed. Twenty-five of the 26 stones consisted of 90 per cent magnesium ammonium phosphate and 10 per cent carbonate apatite. The remaining single stone was composed of 90 per cent calcium oxalate and 10 per cent magnesium ammonium phosphate. Renal function improved significantly with the removal of the stones.

Radiographic changes of the jawbones in end stage renal disease.

Thirty-eight patients with a history of end stage renal disease (ESRD) were evaluated with panoramic and periapical radiographs for changes in lamina dura, trabecular pattern, radiographic density, and pulp chamber size, as well as the presence of radiolucent lesions not associated with the roots of teeth. Partial or complete loss of lamina dura, delicate of absent trabecular patterns, and an overall granular or chalky white appearance associated with an increase in radiographic density were the most common alterations. These tended to occur together and were most easily observed in the lower molar area superior to the mandibular canal. Radiolucent lesions not associated with the roots of teeth, expansion of alveolar ridges, and radiographic loss of the cortical borders of the maxillary sinus, nasal floor, and mandibular canal were less common but were occasionally noted. Multiple correlation coefficients between radiographic changes and selected biomedical patient data were obtained by computer analysis, and their significance discussed. The pathogenesis of end stage renal disease was also reviewed.