A comparative study of the therapeutic effect of bone marrow mesenchymal stem cells versus insulin on mandibular dento-alveolar complex collagen formation and beta-catenin expression in experimentally induced type I diabetes.

Objective: To assess and compare the therapeutic effect of bone marrow mesenchymal stem cells (BM-MSCs) versus insulin on mandibular dento-alveolar complex collagen formation and beta-catenin (ß-catenin) expression in experimentally induced type I diabetes in albino rat. Design: Twenty-eight male albino rats were equally divided as follows; Group I: was composed of rats which received no drug. The remaining rats were administrated a single streptozotocin (STZ) (40 mg/kg) intra-peritoneal injection. After affirmation of diabetes induction, the rats were divided into: Group II: Diabetic rats were given no treatment. Group III: Diabetic rats received a single BM-MSCs intravenous injection (1x106 cells). Group IV: Diabetic rats were given a daily insulin subcutaneous injection (5 IU/kg). After 28 days, mandibles were processed and stained by Hematoxylin & Eosin (H&E), Masson's trichrome and anti-ß-catenin antibody. A statistical analysis was performed to measure positive area% of Masson's trichrome and ß-catenin. Results: Dento-alveolar complex tissues and cells of Group II showed destructive changes histologically, while Groups III and IV demonstrated improved histological features. Group II presented almost old collagen in all dento-alveolar complex tissues, and nearly negative ß-catenin expression. Groups III and IV revealed a newly formed collagen intermingled with very few areas of old collagen, and both groups showed positive ß-catenin immunoreactivity. Statistically, Groups III and IV represented the highest mean values of Masson's trichrome area% and ß-catenin area%, while Group II reported the lowest mean. Conclusions: Streptozotocin has a destructive effect on the dento-alveolar complex structure and function. BM-MSCs and insulin show regenerative capacity in STZ-affected periodontal tissues, and statistically, they increase collagen formation and ß-catenin expression.

Reparative potential of mesenchymal stem cells and platelet-rich plasma on irradiated submandibular glands of male albino rats.

OBJECTIVE: To appraise and compare the reparative role of bone marrow-mesenchymal stem cells (BM-MSCs) and platelet-rich plasma (PRP) against irradiation damage on albino rats' submandibular gland. DESIGN: Seventy four male albino rats were used, one for BM-MSCs harvesting, 10 for PRP preparation, seven as control group (Group 1). The remaining 56 rats were subjected to single dose (6 Gy) gamma irradiation and were divided into equal four groups; (Group 2): received no treatment, (Group 3): each rat was injected with 1 × 105 BM-MSCs, (Group 4): each rat was injected with 0.5 ml/kg PRP, and (Group 5): each rat was injected with 1 × 105 BM-MSCs and 0.5 ml/kg PRP. Each group was further subdivided into two subgroups in which rats sacrificed after one and two weeks from irradiation. Any structural changes were examined histopathologically, immunohistochemically using proliferating cell nuclear antigen (PCNA) and CD31 primary antibodies and histochemically using picrosirius red (PSR) stain, then analyzed statistically. RESULTS: Histopathological examination of Group 2 showed atrophied acini, with nuclear changes and signs of degeneration in duct systems. Treated groups revealed signs of regeneration in form of uniform acini and regenerated duct systems especially in Group 5 and in a time depended manner. Immunohistochemical examination revealed increased immunoexpression of PCNA and CD31, while histochemical examination showed decreased PSR in all treated groups in relation to the irradiated group and this was proved statistically. CONCLUSIONS: BM-MSCs and PRP are effective as treatment for irradiation-induced submandibular gland damage. However, the combined therapy is recommended over each one separately.

Histological, immunohistochemical and radiographic evaluation of Amitriptyline administration on the periodontium of albino rats (An experimental study).

Background: Amitriptyline is a tricyclic antidepressant drug accustomed to treat depressive disorders. It recorded many side effects on different tissues. Objective: To investigate reaction of Albino rats' periodontium after oral administration of Amitriptyline histologically and radiographically. Methods: Fourteen adult male albino rats (150-200 g) were divided into two groups, control and experimental. Rats of experimental group received 10 mg/kg/day of Amitriptyline hydrochloride by oral gavage for four weeks. Mandibles were prepared for hematoxylin and eosin (H&E) and anti-osteopontin (Anti-OPN) immunohistochemistry staining. Bone mineral density was measured in mandibular alveolar bone. Statistical analysis for Anti-OPN and relative Hounsfield unit value (HU value) was performed using independent-samples t-test. Results: Gingiva of experimental group showed epithelial degeneration with pyknotic nuclei and disintegration in lamina propria. Areas of separation in alveolar bone and degeneration of some regions in cementum were seen with apparent increase in periodontal ligament (PDL) thickness and its detachment from bone and cementum at some regions. Immunohistochemical examination of experimental group showed apparently increased immunopositivity in gingiva, cementocytes, osteocytes, cementum, bone matrices, fibroblasts and PDL fibers when compared to control group. Statistical analysis revealed insignificant difference of Anti-OPN area% in gingiva between both studied groups. While there was statistical significant increase of Anti-OPN area% in the other periodontium tissues and high statistical significant decrease of relative HU value in experimental group when compared to control. Conclusions: Amitriptyline has destructive effect on periodontal tissues and statistically increases the expression of Anti-OPN in all periodontal tissues except gingiva and decreases bone mineral density.

Assessment of ultra-structure, viability and function of lipopolysaccharides-stimulated human dermal fibroblasts treated with chrysin and exosomes (in vitro study).

Background: Lipopolysaccharides (LPS) stimulate production of inflammatory cytokines. Chrysin is flavonoid beneficial for treatment of inflammatory conditions. Bone marrow mesenchymal stem cell (BM-MSC) exosomes have regenerative ability in different tissues. Objective: To assess potential role of chrysin and BM-MSC exosomes on ultra-structure, viability and function of human dermal fibroblasts-adult (HDFa) stimulated by LPS. Methods: HDFa cells were divided into: Group I: Cells received no treatment. Group II: Cells were stimulated with LPS. Group III: LPS stimulated cells were treated with chrysin. Group IV: LPS stimulated cells were treated with exosomes. Results: After 48 h, ultrastructural examination of HDFa cells in Group I revealed intact plasma membrane and numerous cytoplasmic organelles. Group II displayed destructed plasma membrane and apoptotic bodies. Group III showed intact plasma membrane with loss of its integrity at some areas. Group IV demonstrated intact plasma membrane that showed fusion with exosomes at some areas. Statistical analysis of MTT represented highest mean value of cell viability% in Group IV followed by Groups III, I and II respectively. Statistical analysis of enzyme-linked immunosorbent assay (ELISA) showed the highest mean value of interleukin-1ß (IL-1ß) was in Group II followed by Groups III, IV and I, while highest mean values of interleukin-10 (IL-10), nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins were in Group I, followed by Groups IV, III and II respectively. Conclusions: LPS have harmful consequences on ultra-structure, viability and function of HDFa cells. BM-MSC exosomes have better regenerative action on inflamed fibroblasts in comparison to chrysin.

Regenerative capacity of bone marrow stem cells on aged albino rat's parotid excretory duct.

OBJECTIVE: To appraise structural features of parotid excretory duct during senility and probable effect of bone marrow-mesenchymal stem cells (BM-MSCs). DESIGN: A total of 14 healthy male albino rats were used. Seven adult rats (24-34-week-old) represented the control group (Group I). Seven senile rats (72-80-week-old) were utilized in which the left parotid gland served as "Old" (Group II) and were injected by 0.2 ml phosphate buffered saline; the right side represented "Old treated" (Group III) and got local injection of 1-1.5 million allogeneic BM-MSCs. One month later, glands were dissected and assessed structurally, ultra-structurally and statistically. RESULT: Histologically, Group I showed normal duct histology. In Group II duct lining lost its pseudostratification which was recovered in Group III. PCNA immunolocalization showed moderate reactivity in Group I, negative to mild reaction in Group II, and strong reaction in some of Group III cells. Ultra-structural features of Group I were ordinary in which basal cell had a large flat nucleus, and dark and light cells showed electron-dense cytoplasm and electron-lucent cytoplasm respectively. Tuft cell displayed long microvilli. Mucous droplets filled goblet cell. Group II revealed an apparent reduction in cells size, organelles and absence of tuft cell. In Group III all cell types were detected and they recovered their organelles, cell and nucleus shape. The highest mean area% of PCNA immunoreactivity was in Group I followed by Group III then Group II. CONCLUSIONS: Aging has a deteriorating effect on structure and ultra-structure of parotid gland excretory duct that could be amended by BM-MSCs.

The prospective role of mesenchymal stem cells exosomes on circumvallate taste buds in induced Alzheimer's disease of ovariectomized albino rats: (Light and transmission electron microscopic study).

OBJECTIVE: To elucidate the effect of Alzheimer's disease on the structure of circumvallate papilla taste buds and the possible role of exosomes on the taste buds in Alzheimer's disease. DESIGN: Forty two ovariectomized female adult albino rats were utilized and divided into: Group I: received vehicle. Group II: received aluminum chloride to induce Alzheimer's disease. Group III: after the induction of Alzheimer's disease, each rat received single dose of exosomes then left for 4 weeks. The circumvallate papillae were prepared for examination by light and transmission electron microscope. STATISTICAL ANALYSIS: histomorphometric data were statistically analyzed. RESULTS: Histological examination of circumvallate papilla in Group I showed normal histological features. Group II revealed distorted features. Group III illustrated nearly normal histological features of circumvallate. Silver impregnation results showed apparently great number of heavily impregnated glossopharyngeal nerve fibers in both Groups I & III but markedly decreased in Group II. Synaptophysin-immunoreactivity was strong in Group I, mild in Group II and moderate in Group III. The ultra-structural examination of taste bud cells revealed normal features in Group I, distorted features in Group II and almost normal features in Group III. Statistically highest mean of Synaptophysin-immunoreactivity area% was for Group I, followed by Group III, and the least value was for Group II. CONCLUSIONS: Alzheimer's disease has degenerative effects. Bone marrow mesenchymal stem cell (BM-MSC)-derived exosomes have the ability to improve the destructive changes induced by Alzheimer's disease.

Role of bone marrow-derived stem cells versus insulin on filiform and fungiform papillae of diabetic albino rats (light, fluorescent and scanning electron microscopic study).

BACKGROUND: Diabetes mellitus (DM) is a chronic metabolic disease characterized by high blood glucose levels. DM affects many body's organs and caused by insulin production deficiency or by the ineffectiveness of the produced insulin. Administration of exogenous insulin is required for management of type I DM; however, it does not cure the disease. Bone marrow-mesenchymal stem cells (BM-MSCs) have been highlighted to offer a novel cell based approach for treatment of diabetes because of their anti-diabetic effect, direct differentiation into a variety of cell types, or release of paracrine factors. AIM: To examine the effect of BM-MSCs versus insulin on true filiform and fungiform papillae of diabetic rats. MATERIALS AND METHODS: Fifty six male Wistar albino rats weighing 200-250 g were equally divided into: Control group (Gp I): Rats did not receive any drug. Diabetic group (Gp II): Rats received a single intra-peritoneal injection of streptozotocin (40 mg/kg). BM-MSCs treated diabetic group (Gp III): After DM confirmation; rats received a single intravenous injection of BM-MSCs (million units) through tail vein. Insulin treated diabetic group (Gp IV): After DM confirmation; rats received a daily subcutaneous injection of insulin (5IU/kg). After four weeks, half of the tongue specimens were processed and stained by Hematoxyline & Eosin and Anti-proliferating cell nuclear antigen (Anti-PCNA) then examined by light microscope. Fluorescent microscope was used to detect homing of injected labeled BM-MSCs in rats' filiform and fungiform papillae. While the other half were examined by scanning electron microscope. RESULTS: True filiform and fungiform papillae of Gp II showed significant histological and morphological alterations. In treated groups, Gp III and Gp IV, both papillae showed marked improvements, being more noticeable in Gp IV. There was a significant increase in the number of Anti-PCNA positive cells and a significant decrease in fasting blood glucose level in Gp III and Gp IV in comparison to Gp II. CONCLUSIONS: DM had degenerative effects on true filiform and fungiform papillae. Administration of BM-MSCs reduced the deleterious effects of DM on both papillae. Insulin injection caused more obvious improvements in both papillae of diabetic rats than BM-MSCs.

Intravenous patient-controlled fentanyl with and without transversus abdominis plane block in cirrhotic patients post liver resection.

BACKGROUND: Coagulation changes can complicate liver resection, particularly in patients with cirrhosis. The aim of this prospective hospital-based comparative study was to compare the postoperative analgesic efficacy of intravenous fentanyl patient-controlled analgesia (IVPCA) with and without transversus abdominis plane (TAP) block. METHODS: Fifty patients with Child's A cirrhosis undergoing liver resection were randomly divided into two groups for postoperative analgesia, ie, an IVPCA group receiving a 10 µg/mL fentanyl bolus of 15 µg with a 10-minute lockout and a maximum hourly dose of 90 µg, and an IVPCA + TAP group that additionally received TAP block (15 mL of 0.375% bupivacaine) on both sides via a posterior approach with ultrasound guidance before skin incision. Postoperatively, bolus injections of bupivacaine 0.375% were given every 8 hours through a TAP catheter inserted by the surgeon in the open space during closure of the inverted L-shaped right subcostal with midline extension (subcostal approach) guided by the visual analog scale score (<3, 5 mL; 3 to <6, 10 mL; 6-10, 15-20 mL) according to weight (maximum 2 mg/kg). The top-up dosage of local anesthetic could be omitted if the patient was not in pain. Coagulation was monitored using standard coagulation tests. RESULTS: Age, weight, and sex were comparable between the groups (P>0.05). The visual analog scale score was significantly lower at 12, 18, 24, 48, and 72 hours (P<0.01) in IVPCA + TAP group. The Ramsay sedation score was lower only after 72 hours in the IVPCA + TAP group when compared with the IVPCA group (1.57±0.74 versus 2.2±0.41, respectively, P<0.01). Heart rate, systolic blood pressure, and fentanyl consumption were lower in the IVPCA + TAP group at 24, 48, and 72 hours (P<0.05). Intensive care unit stays were significantly shorter with TAP (2.61±0.74 days versus 4.35±0.79 days, P<0.01). Prothrombin time and International Normalized Ratio indicated temporary hypocoagulability in both groups. CONCLUSION: Combining TAP with IVPCA improved postoperative pain management and reduced fentanyl consumption, with a shorter stay in intensive care. TAP block can be included as part of a balanced multimodal postoperative pain regimen.