Effect of Stem Cells, Ascorbic Acid and SERCA1a Gene Transfected Stem Cells in Experimentally Induced Type I Diabetic Myopathy

Background and Objectives@#Sarco/endoplasmic reticulum Ca2＋-ATPase (SERCA) inhibition was proved in streptozotocin (STZ)-diabetic rats. The present study aimed at investigating and comparing the therapeutic effect of bone marrow mesenchymal stem cells (BMMSCs), BMMSCs combined with ascorbic acid (AA) and SERCA1a gene transfected BMMSCs in induced type I diabetic myopathy of male albino rat. @*Methods@#and Results: 54 rats were divided into donor group of 6 rats for isolation, propagation and characterization of BMMSCs and SERCA1a transfected BMMSCs, groups I∼V 48 rats. Group I of 8 control rats, group II (Diabetic) of 10 rats given STZ 50 mg/kg intraperitoneal, group III (BMMSCs) of 10 rats given STZ and BMMSCs intravenous (IV), group IV (BMMSCs and AA) of 10 rats given STZ, BMMSCs IV and AA 500 mg/kg and group V (SERCA 1a transfected BMMSCs) of 10 rats given STZ and SERCA1a transfected BMMSCs IV. The rats were sacrificed after 8 weeks. Gastrocnemius specimens were subjected to biochemical, histological, morphometric and statistical studies. Diabetic rats revealed inflammatory and degenerative muscle changes, a significant increase in blood glucose level, mean DNA fragmentation and mean MDA values and a significant decrease in mean GSH and catalase values, area of pale nuclei, area% of CD105 and CD34 ＋ve cells, SERCA1a protein and gene values. The morphological changes regressed by therapy. In group III significant decrease in DNA fragmentation and MDA, significant increase in GSH and catalase, significant increase in the mean area of pale nuclei, area % of CD105 and CD34 ＋ve cells versus diabetic group. In group IV, same findings as group III versus diabetic and BMMSCs groups. In group V, same findings as group IV versus diabetic and treated groups. Western blot and PCR proved a mean value of SERCA1a protein and gene comparable to the control group. Mean calcium concentration values revealed a significant increase in the diabetic group, in BMMSCs and AA group versus control and SERCA1a group. @*Conclusions@#SERCA1a transfected BMMSCs proved a definite therapeutic effect, more remarkable than BMMSCs combined with AA. This effect was evidenced histologically and confirmed by significant changes in the biochemical tests indicating oxidative stress, muscle calcium concentration, morphometric parameters and PCR values of SERCA1a.

effect of microcurrent therapy on repair of induced tendon injury in albino rat: possible role of endogenous stem cells

Tendon repair involves a slow repair process, which results in inferior repair of tissue and failure to obtain full active range of motion. Microcurrent therapy [MCT] is a new therapy after arthroplasty. Stem cells with capacity of self-renewal are ideal for tissue engineering. The present work aimed at investigating the effect of MCT and the possible role of endogenous stem cells in repair of induced tendon injury in albino rat. Twenty-four male albino rats were classified into: Control group, tenotomy group, 5 rats were sacrificed 2 weeks and 5 rats sacrificed 4 weeks following achillis tendon injury [Subgroups IIa and IIb respectively]. MCT group of 10 rats subjected to tendon injury followed by MCT, 5 rats sacrificed 2 weeks and 5 rats sacrificed 4 weeks following MCT [Subgroups IIIa and IIIb respectively]. Tendon sections were stained with H and E and CD44 and CD105 immunostains. A morphometric study was performed. Subgroup IIa demonstrated multiple areas of widely separated collagen fibers, infiltrating cells and multiple congested vessels. Subgroup IIb showed multiple areas of disorganized collagen bundles, less infiltrating cells and few congested vessels. MCT subgroup IIIa revealed some areas of parallel collagen bundles, some infiltrating cells and some dilated congested vessels. MCT subgroup IIIb showed multiple areas of parallel collagen bundles, few infiltrating cells and occasional congested vessels. CD44 +ve and CD105 +ve cells were seen among disorganized, parallel collagen fibers and inside blood vessels. A significant increase in the area of regenerated collagen bundles was recorded in MCT subgroup IIIb. The area% of CD44 +ve and CD105 +ve MSCs denoted a significant increase in MCT subgroup IIIa. MCT activated endogenous bone marrow derived MSCs migration to the injured achillis tendon, which stimulated tendon repair following induced tenotomy

Relation between microcurrent therapy and satellite cells in the regeneration of induced skeletal muscle injury in rat

Severe injuries in skeletal muscle result in muscle weakness, which delays recovery and contributes to progressive decline in muscle function. Microcurrent therapy is a novel treatment method used in soft-tissue injury and tissue regeneration therapy. The regenerative capacity of skeletal muscle tissue resides in satellite cells, the quiescent adult stem cells. The present work aimed at investigating the possible relation between microcurrent therapy and satellite cells in regeneration of induced skeletal muscle injury in albino rats. Twenty-four male albino rats were divided into 2 groups: Control group and experimental group [II]: rats were subjected to gastrocnemius-soleus muscle injury [subgroup IIa], they were subdivided into subgroups IIa1 and IIa2 sacrificed 1 and 3 weeks after injury respectively. Subgroup IIb: Rats were subjected to muscle injury and micro-current electric stimulator, was applied for 20 minutes for three sessions per week. The animals were subdivided into subgroups IIb1 and IIb2 sacrificed 1 and 3 weeks following the day of injury. Muscle sections were stained with hematoxylin and eosin, alpha smooth muscle actin [alpha-SMA] and CD34 immunostaining. Morphometric studies and statistical analysis were performed. Atypical fibers were widely separated by connective tissue cells and revealed partial loss of striations in subgroup IIa. Some fibers recruited strong acidophilic sarcoplasm with focal vacuolations in subgroup IIa1. In subgroup IIb1, some typical fibers, some centrally located nuclei, and a few deeply acidophilic fibers were found. Striations were found in some areas of the sarcoplasm. In subgroup IIb2 striations were found in most areas of the sarcoplasm. A significant decrease in the mean area of atypical fibers, a significant increase in the mean area% of alpha-smooth muscle actin-positive cells, and a significant increase in the mean area% of CD34-positive cells were found in subgroup IIb compared with subgroup IIa. A definite therapeutic effect of the microcurrent was found on induced skeletal muscle injury, which was time dependent. This effect was proved to be related to satellite cell activation